Inhibitors of lysine specific demethylase-1

ABSTRACT

The present invention relates generally to compositions and methods for treating cancer and neoplastic disease. Provided herein are substituted heterocyclic derivative compounds and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for inhibition of lysine specific demethylase-1. Furthermore, the subject compounds and compositions are useful for the treatment of cancer.

CROSS REFERENCE

This application is a continuation of U.S. application Ser. No.15/401,894, filed Jan. 9, 2017, which is a continuation of Ser. No.14/988,022, filed Jan. 5, 2016, now U.S. Pat. No. 9,573,930, which is acontinuation of Ser. No. 14/701,304, filed Apr. 30, 2015, now U.S. Pat.No. 9,255,097, which claims the priority benefit of U.S. ProvisionalApplication No. 61/987,354, filed May 1, 2014, the contents of which arehereby incorporated by reference in their entireties for all purposes.

BACKGROUND

A need exists in the art for an effective treatment of cancer andneoplastic disease.

BRIEF SUMMARY OF THE INVENTION

Provided herein are substituted heterocyclic derivative compounds andpharmaceutical compositions comprising said compounds. The subjectcompounds and compositions are useful for inhibition lysine specificdemethylase-1 (LSD-1). Furthermore, the subject compounds andcompositions are useful for the treatment of cancer, such as acutemyeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small celllung cancer (SCLC), non-small cell lung cancer (NSCLC), neuroblastoma,small round blue cell tumors, glioblastoma, prostate cancer, breastcancer, bladder cancer, lung cancer and/or melanoma and the like. Thesubstituted heterocyclic derivative compounds described herein are basedupon a central heterocyclic ring system, such as pyrimidinone, or thelike. Said pyrimidinone ring system is further substituted with a4-cyanophenyl group and additional groups, such as aryl, heteroaryl orheterocyclic groups.

One embodiment provides a compound having the structure of Formula (I),or a pharmaceutically acceptable salt thereof,

wherein,

W is N, C—H, or C—F;

X is hydrogen, halogen, —CN, optionally substituted alkyl, optionallysubstituted alkynyl, optionally substituted carbocyclylalkynyl,optionally substituted aryl, or optionally substituted heteroaryl;

Y is hydrogen, optionally substituted alkyl, optionally substitutedcycloalkyl, or optionally substituted cycloalkylalkyl;

Z is an optionally substituted group chosen from alkyl, carbocyclyl,C-attached heterocyclyl, N-attached heterocyclyl, heterocyclylalkyl,heterocyclylalkenyl, —O-heterocyclyl, —N(R)— heterocyclyl,—O-heterocyclylalkyl, —N(R)-heterocyclylalkyl, —N(R)(C₁-C₄alkylene)-NR₂,—O(C₁-C₄alkylene)-NR₂, and R is hydrogen or C₁-C₄alkyl.

One embodiment provides a compound having the structure of Formula (Ia),or a pharmaceutically acceptable salt thereof,

wherein,

W is N, C—H, or C—F;

X is hydrogen, halogen, —CN, optionally substituted alkynyl, optionallysubstituted carbocyclylalkynyl, optionally substituted aryl, oroptionally substituted heteroaryl;

Y is hydrogen, optionally substituted alkyl, optionally substitutedcycloalkyl, or optionally substituted cycloalkylalkyl; and

Z is an optionally substituted group chosen from N-attachedheterocyclyl, —O— heterocyclylalkyl, —N(H)-heterocyclyl,—N(Me)-heterocyclyl, —N(H)-heterocyclylalkyl, or—N(Me)-heterocyclylalkyl.

One embodiment provides a compound having the structure of Formula (Ib),or a pharmaceutically acceptable salt thereof,

wherein,

-   -   W is N, C—H, or C—F;    -   X is hydrogen, halogen, optionally substituted alkynyl,        optionally substituted carbocyclylalkynyl, optionally        substituted aryl, or optionally substituted heteroaryl;    -   Y is hydrogen, optionally substituted alkyl, or optionally        substituted cycloalkyl; and    -   Z is an optionally substituted group chosen from N-heterocyclyl,        —O— heterocyclylalkyl, —N(H)-heterocyclylalkyl, or        —N(Me)-heterocyclylalkyl.

One embodiment provides a pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient. One embodiment provides apharmaceutical composition comprising a compound of Formula (Ia), or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient. One embodiment provides a pharmaceuticalcomposition comprising a compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable excipient.

One embodiment provides a method of regulating gene transcription in acell comprising inhibiting lysine-specific demethylase 1 activity byexposing the lysine-specific demethylase 1 enzyme to a compound ofFormula (I). One embodiment provides a method of regulating genetranscription in a cell comprising inhibiting lysine-specificdemethylase 1 activity by exposing the lysine-specific demethylase 1enzyme to a compound of Formula (Ia). One embodiment provides a methodof regulating gene transcription in a cell comprising inhibitinglysine-specific demethylase 1 activity by exposing the lysine-specificdemethylase 1 enzyme to a compound of Formula (Ib).

One embodiment provides a method of treating cancer in a patient in needthereof, comprising administering to the patient a compound of Formula(I), or a pharmaceutically acceptable salt thereof. One embodimentprovides a method of treating cancer in a patient in need thereof,comprising administering to the patient a compound of Formula (Ia), or apharmaceutically acceptable salt thereof. One embodiment provides amethod of treating cancer in a patient in need thereof, comprisingadministering to the patient a compound of Formula (Ib), or apharmaceutically acceptable salt thereof.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference for the specificpurposes identified herein.

DETAILED DESCRIPTION OF THE INVENTION

As used herein and in the appended claims, the singular forms “a,”“and,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “an agent” includesa plurality of such agents, and reference to “the cell” includesreference to one or more cells (or to a plurality of cells) andequivalents thereof known to those skilled in the art, and so forth.When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange, in some instances, will vary between 1% and 15% of the statednumber or numerical range. The term “comprising” (and related terms suchas “comprise” or “comprises” or “having” or “including”) is not intendedto exclude that in other certain embodiments, for example, an embodimentof any composition of matter, composition, method, or process, or thelike, described herein, “consist of” or “consist essentially of” thedescribed features.

Definitions

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated below.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Thioxo” refers to the ═S radical.

“Imino” refers to the ═N—H radical.

“Oximo” refers to the ═N—OH radical.

“Hydrazino” refers to the ═N—NH₂ radical.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to fifteen carbon atoms (e.g., C₁-C₁₅alkyl). In certain embodiments, an alkyl comprises one to thirteencarbon atoms (e.g., C₁-C₁₃ alkyl). In certain embodiments, an alkylcomprises one to eight carbon atoms (e.g., C₁-C₈ alkyl). In otherembodiments, an alkyl comprises one to five carbon atoms (e.g., C₁-C₅alkyl). In other embodiments, an alkyl comprises one to four carbonatoms (e.g., C₁-C₄ alkyl). In other embodiments, an alkyl comprises oneto three carbon atoms (e.g., C₁-C₃ alkyl). In other embodiments, analkyl comprises one to two carbon atoms (e.g., C₁-C₂ alkyl). In otherembodiments, an alkyl comprises one carbon atom (e.g., C₁ alkyl). Inother embodiments, an alkyl comprises five to fifteen carbon atoms(e.g., C₅-C₁₅ alkyl). In other embodiments, an alkyl comprises five toeight carbon atoms (e.g., C₅-C₈ alkyl). In other embodiments, an alkylcomprises two to five carbon atoms (e.g., C₂-C₅ alkyl). In otherembodiments, an alkyl comprises three to five carbon atoms (e.g., C₃-C₅alkyl). In other embodiments, the alkyl group is selected from methyl,ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl(n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl),1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl isattached to the rest of the molecule by a single bond. Unless statedotherwise specifically in the specification, an alkyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)—N(R^(a))₂,—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2)and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkoxy” refers to a radical bonded through an oxygen atom of theformula —O-alkyl, where alkyl is an alkyl chain as defined above.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon double bond, and having from two to twelvecarbon atoms. In certain embodiments, an alkenyl comprises two to eightcarbon atoms. In other embodiments, an alkenyl comprises two to fourcarbon atoms. The alkenyl is attached to the rest of the molecule by asingle bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e.,allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unlessstated otherwise specifically in the specification, an alkenyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)— N(R^(a))₂,—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2)and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon triple bond, having from two to twelve carbonatoms. In certain embodiments, an alkynyl comprises two to eight carbonatoms. In other embodiments, an alkynyl comprises two to six carbonatoms. In other embodiments, an alkynyl comprises two to four carbonatoms. The alkynyl is attached to the rest of the molecule by a singlebond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, andthe like. Unless stated otherwise specifically in the specification, analkynyl group is optionally substituted by one or more of the followingsubstituents: halo, cyano, nitro, oxo, thioxo, imino, oximo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)—N(R^(a))₂, —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or2), —S(O)_(t)OR^(a)(where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, containing no unsaturation andhaving from one to twelve carbon atoms, for example, methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain isattached to the rest of the molecule through a single bond and to theradical group through a single bond. The point(s) of attachment of thealkylene chain to the rest of the molecule and to the radical group isthrough one carbon in the alkylene chain or through any two carbonswithin the chain. In certain embodiments, an alkylene comprises one toeight carbon atoms (e.g., C₁-C₈ alkylene). In other embodiments, analkylene comprises one to five carbon atoms (e.g., C₁-C₅ alkylene). Inother embodiments, an alkylene comprises one to four carbon atoms (e.g.,C₁-C₄ alkylene). In other embodiments, an alkylene comprises one tothree carbon atoms (e.g., C₁-C₃ alkylene). In other embodiments, analkylene comprises one to two carbon atoms (e.g., C₁-C₂ alkylene). Inother embodiments, an alkylene comprises one carbon atom (e.g., C₁alkylene). In other embodiments, an alkylene comprises five to eightcarbon atoms (e.g., C₅-C₈ alkylene). In other embodiments, an alkylenecomprises two to five carbon atoms (e.g., C₂-C₅ alkylene). In otherembodiments, an alkylene comprises three to five carbon atoms (e.g.,C₃-C₅ alkylene). Unless stated otherwise specifically in thespecification, an alkylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, oxo, thioxo,imino, oximo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —OC(O)— N(R^(a))₂, —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), aryl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), aralkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclylalkyl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), heteroaryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl).

“Alkynylene” or “alkynylene chain” refers to a straight or brancheddivalent hydrocarbon chain linking the rest of the molecule to a radicalgroup, consisting solely of carbon and hydrogen, containing at least onecarbon-carbon triple bond, and having from two to twelve carbon atoms.The alkynylene chain is attached to the rest of the molecule through asingle bond and to the radical group through a single bond. In certainembodiments, an alkynylene comprises two to eight carbon atoms (e.g.,C₂-C₈ alkynylene). In other embodiments, an alkynylene comprises two tofive carbon atoms (e.g., C₂-C₅ alkynylene). In other embodiments, analkynylene comprises two to four carbon atoms (e.g., C₂-C₄ alkynylene).In other embodiments, an alkynylene comprises two to three carbon atoms(e.g., C₂-C₃ alkynylene). In other embodiments, an alkynylene comprisestwo carbon atom (e.g., C₂ alkylene). In other embodiments, an alkynylenecomprises five to eight carbon atoms (e.g., C₅-C₈ alkynylene). In otherembodiments, an alkynylene comprises three to five carbon atoms (e.g.,C₃-C₅ alkynylene). Unless stated otherwise specifically in thespecification, an alkynylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, oxo, thioxo,imino, oximo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —OC(O)— N(R^(a))₂, —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), aryl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), aralkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclylalkyl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), heteroaryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl).

“Aryl” refers to a radical derived from an aromatic monocyclic ormulticyclic hydrocarbon ring system by removing a hydrogen atom from aring carbon atom. The aromatic monocyclic or multicyclic hydrocarbonring system contains only hydrogen and carbon from five to eighteencarbon atoms, where at least one of the rings in the ring system isfully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)it-electron system in accordance with the Hückel theory. The ring systemfrom which aryl groups are derived include, but are not limited to,groups such as benzene, fluorene, indane, indene, tetralin andnaphthalene. Unless stated otherwise specifically in the specification,the term “aryl” or the prefix “ar-” (such as in “aralkyl”) is meant toinclude aryl radicals optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O) R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Aralkyl” refers to a radical of the formula —R^(c)-aryl where R^(c) isan alkylene chain as defined above, for example, methylene, ethylene,and the like. The alkylene chain part of the aralkyl radical isoptionally substituted as described above for an alkylene chain. Thearyl part of the aralkyl radical is optionally substituted as describedabove for an aryl group.

“Aralkenyl” refers to a radical of the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula —R^(e)-aryl, where R^(e)is an alkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Aralkoxy” refers to a radical bonded through an oxygen atom of theformula —O—R^(c)-aryl where R^(c) is an alkylene chain as defined above,for example, methylene, ethylene, and the like. The alkylene chain partof the aralkyl radical is optionally substituted as described above foran alkylene chain. The aryl part of the aralkyl radical is optionallysubstituted as described above for an aryl group.

“Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which includes fused or bridged ring systems, having from three tofifteen carbon atoms. In certain embodiments, a carbocyclyl comprisesthree to ten carbon atoms. In other embodiments, a carbocyclyl comprisesfive to seven carbon atoms. The carbocyclyl is attached to the rest ofthe molecule by a single bond. Carbocyclyl is saturated (i.e.,containing single C—C bonds only) or unsaturated (i.e., containing oneor more double bonds or triple bonds). A fully saturated carbocyclylradical is also referred to as “cycloalkyl.” Examples of monocycliccycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl isalso referred to as “cycloalkenyl.” Examples of monocyclic cycloalkenylsinclude, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, andcyclooctenyl. Polycyclic carbocyclyl radicals include, for example,adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl,decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unlessotherwise stated specifically in the specification, the term“carbocyclyl” is meant to include carbocyclyl radicals that areoptionally substituted by one or more substituents independentlyselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Carbocyclylalkyl” refers to a radical of the formula —R^(c)-carbocyclylwhere R^(c) is an alkylene chain as defined above. The alkylene chainand the carbocyclyl radical is optionally substituted as defined above.

“Carbocyclylalkynyl” refers to a radical of the formula—R^(c)-carbocyclyl where R^(c) is an alkynylene chain as defined above.The alkynylene chain and the carbocyclyl radical is optionallysubstituted as defined above.

“Carbocyclylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-carbocyclyl where R^(c) is an alkylene chain asdefined above. The alkylene chain and the carbocyclyl radical isoptionally substituted as defined above.

As used herein, “carboxylic acid bioisostere” refers to a functionalgroup or moiety that exhibits similar physical, biological and/orchemical properties as a carboxylic acid moiety. Examples of carboxylicacid bioisosteres include, but are not limited to,

and the like.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodosubstituents.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, fluoromethyl,2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. Insome embodiments, the alkyl part of the fluoroalkyl radical isoptionally substituted as defined above for an alkyl group.

“Heterocyclyl” refers to a stable 3- to 18-membered non-aromatic ringradical that comprises two to twelve carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radical isa monocyclic, bicyclic, tricyclic or tetracyclic ring system, whichoptionally includes fused or bridged ring systems. The heteroatoms inthe heterocyclyl radical are optionally oxidized. One or more nitrogenatoms, if present, are optionally quaternized. The heterocyclyl radicalis partially or fully saturated. The heterocyclyl is attached to therest of the molecule through any atom of the ring(s). Examples of suchheterocyclyl radicals include, but are not limited to, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, the term “heterocyclyl” is meant to include heterocyclylradicals as defined above that are optionally substituted by one or moresubstituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,oxo, thioxo, cyano, nitro, optionally substituted aryl, optionallysubstituted aralkyl, optionally substituted aralkenyl, optionallysubstituted aralkynyl, optionally substituted carbocyclyl, optionallysubstituted carbocyclylalkyl, optionally substituted heterocyclyl,optionally substituted heterocyclylalkyl, optionally substitutedheteroaryl, optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“N-heterocyclyl” or “N-attached heterocyclyl” refers to a heterocyclylradical, as defined above, containing at least one nitrogen and wherethe point of attachment of the heterocyclyl radical to the rest of themolecule is through a nitrogen atom in the heterocyclyl radical. AnN-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such N-heterocyclyl radicals include,but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl,1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

“C-heterocyclyl” or “C-attached heterocyclyl” refers to a heterocyclylradical as defined above containing at least one heteroatom and wherethe point of attachment of the heterocyclyl radical to the rest of themolecule is through a carbon atom in the heterocyclyl radical. AC-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such C-heterocyclyl radicals include,but are not limited to, 2-morpholinyl, 2- or 3- or 4-piperidinyl,2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.If the heterocyclyl is a nitrogen-containing heterocyclyl, theheterocyclyl is optionally attached to the alkyl radical at the nitrogenatom. The alkylene chain of the heterocyclylalkyl radical is optionallysubstituted as defined above for an alkylene chain. The heterocyclylpart of the heterocyclylalkyl radical is optionally substituted asdefined above for a heterocyclyl group.

“Heterocyclylalkoxy” refers to a radical bonded through an oxygen atomof the formula —O—R^(c)-heterocyclyl where R^(c) is an alkylene chain asdefined above. If the heterocyclyl is a nitrogen-containingheterocyclyl, the heterocyclyl is optionally attached to the alkylradical at the nitrogen atom. The alkylene chain of theheterocyclylalkoxy radical is optionally substituted as defined abovefor an alkylene chain. The heterocyclyl part of the heterocyclylalkoxyradical is optionally substituted as defined above for a heterocyclylgroup.

“Heteroaryl” refers to a radical derived from a 3- to 18-memberedaromatic ring radical that comprises two to seventeen carbon atoms andfrom one to six heteroatoms selected from nitrogen, oxygen and sulfur.As used herein, the heteroaryl radical is a monocyclic, bicyclic,tricyclic or tetracyclic ring system, wherein at least one of the ringsin the ring system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) it-electron system in accordance with the Hückeltheory. Heteroaryl includes fused or bridged ring systems. Theheteroatom(s) in the heteroaryl radical is optionally oxidized. One ormore nitrogen atoms, if present, are optionally quaternized. Theheteroaryl is attached to the rest of the molecule through any atom ofthe ring(s). Examples of heteroaryls include, but are not limited to,azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl,benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl,benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl,benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]-pyrimidinyl, pyridinyl,pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetra-hydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, theterm “heteroaryl” is meant to include heteroaryl radicals as definedabove which are optionally substituted by one or more substituentsselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl,haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl,optionally substituted aralkyl, optionally substituted aralkenyl,optionally substituted aralkynyl, optionally substituted carbocyclyl,optionally substituted carbocyclylalkyl, optionally substitutedheterocyclyl, optionally substituted heterocyclylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl,—R^(b)—OR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a),—R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. An N-heteroaryl radical is optionallysubstituted as described above for heteroaryl radicals.

“C-heteroaryl” refers to a heteroaryl radical as defined above and wherethe point of attachment of the heteroaryl radical to the rest of themolecule is through a carbon atom in the heteroaryl radical. AC-heteroaryl radical is optionally substituted as described above forheteroaryl radicals.

“Heteroarylalkyl” refers to a radical of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is optionally attachedto the alkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group.

“Heteroarylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-heteroaryl, where R^(c) is an alkylene chain asdefined above. If the heteroaryl is a nitrogen-containing heteroaryl,the heteroaryl is optionally attached to the alkyl radical at thenitrogen atom. The alkylene chain of the heteroarylalkoxy radical isoptionally substituted as defined above for an alkylene chain. Theheteroaryl part of the heteroarylalkoxy radical is optionallysubstituted as defined above for a heteroaryl group.

The compounds disclosed herein, in some embodiments, contain one or moreasymmetric centers and thus give rise to enantiomers, diastereomers, andother stereoisomeric forms that are defined, in terms of absolutestereochemistry, as (R)- or (S)—. Unless stated otherwise, it isintended that all stereoisomeric forms of the compounds disclosed hereinare contemplated by this disclosure. When the compounds described hereincontain alkene double bonds, and unless specified otherwise, it isintended that this disclosure includes both E and Z geometric isomers(e.g., cis or trans). Likewise, all possible isomers, as well as theirracemic and optically pure forms, and all tautomeric forms are alsointended to be included. The term “geometric isomer” refers to E or Zgeometric isomers (e.g., cis or trans) of an alkene double bond. Theterm “positional isomer” refers to structural isomers around a centralring, such as ortho-, meta-, and para-isomers around a benzene ring.

A “tautomer” refers to a molecule wherein a proton shift from one atomof a molecule to another atom of the same molecule is possible. Thecompounds presented herein, in certain embodiments, exist as tautomers.In circumstances where tautomerization is possible, a chemicalequilibrium of the tautomers will exist. The exact ratio of thetautomers depends on several factors, including physical state,temperature, solvent, and pH. Some examples of tautomeric equilibriuminclude:

“Pharmaceutically acceptable salt” includes both acid and base additionsalts. A pharmaceutically acceptable salt of any one of the substitutedheterocyclic derivative compounds described herein is intended toencompass any and all pharmaceutically suitable salt forms. Preferredpharmaceutically acceptable salts of the compounds described herein arepharmaceutically acceptable acid addition salts and pharmaceuticallyacceptable base addition salts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid,hydrofluoric acid, phosphorous acid, and the like. Also included aresalts that are formed with organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, alkanedioic acids, aromatic acids, aliphatic and. aromaticsulfonic acids, etc. and include, for example, acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Exemplary salts thus include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates,monohydrogen-phosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates,trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates,mandelates, benzoates, chlorobenzoates, methylbenzoates,dinitro-benzoates, phthalates, benzenesulfonates, toluenesulfonates,phenylacetates, citrates, lactates, malates, tartrates,methanesulfonates, and the like. Also contemplated are salts of aminoacids, such as arginates, gluconates, and galacturonates (see, e.g.,Berge S. M. et al., Pharmaceutical Salts, J. Pharma. Sci. 66:1-19(1997)). Acid addition salts of basic compounds are, in someembodiments, prepared by contacting the free base forms with asufficient amount of the desired acid to produce the salt according tomethods and techniques with which a skilled artisan is familiar.

“Pharmaceutically acceptable base addition salt” refers to those saltsthat retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Pharmaceutically acceptable base addition salts are, insome embodiments, formed with metals or amines, such as alkali andalkaline earth metals or organic amines. Salts derived from inorganicbases include, but are not limited to, sodium, potassium, lithium,ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminumsalts and the like. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, for example,isopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine,hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline,N-methylglucamine, glucosamine, methylglucamine, theobromine, purines,piperazine, piperidine, N-ethylpiperidine, polyamine resins and thelike. See Berge et al., supra.

As used herein, “treatment” or “treating,” or “palliating” or“ameliorating” are used interchangeably. These terms refer to anapproach for obtaining beneficial or desired results including but notlimited to therapeutic benefit and/or a prophylactic benefit. By“therapeutic benefit” is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient is still afflicted with the underlying disorder. Forprophylactic benefit, the compositions are, in some embodiments,administered to a patient at risk of developing a particular disease, orto a patient reporting one or more of the physiological symptoms of adisease, even though a diagnosis of this disease has not been made.

“Prodrug” is meant to indicate a compound that is, in some embodiments,converted under physiological conditions or by solvolysis to abiologically active compound described herein. Thus, the term “prodrug”refers to a precursor of a biologically active compound that ispharmaceutically acceptable. A prodrug is typically inactive whenadministered to a subject, but is converted in vivo to an activecompound, for example, by hydrolysis. The prodrug compound often offersadvantages of solubility, tissue compatibility or delayed release in amammalian organism (see, e.g., Bundgard, H., DESIGN OF PRODRUGS (1985),pp. 7-9, 21-24 (Elsevier, Amsterdam).

A discussion of prodrugs is provided in Higuchi, T., et al., Pro-drugsas Novel Delivery Systems, A.C.S. Symposium Series, Vol. 14, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987.

The term “prodrug” is also meant to include any covalently bondedcarriers, which release the active compound in vivo when such prodrug isadministered to a mammalian subject. Prodrugs of an active compound, asdescribed herein, are prepared by modifying functional groups present inthe active compound in such a way that the modifications are cleaved,either in routine manipulation or in vivo, to the parent activecompound. Prodrugs include compounds wherein a hydroxy, amino ormercapto group is bonded to any group that, when the prodrug of theactive compound is administered to a mammalian subject, cleaves to forma free hydroxy, free amino or free mercapto group, respectively.Examples of prodrugs include, but are not limited to, acetate, formateand benzoate derivatives of alcohol or amine functional groups in theactive compounds and the like.

Substituted Heterocyclic Derivative Compounds

Substituted heterocyclic derivative compounds are described herein thatare lysine specific demethylase-1 inhibitors. These compounds, andcompositions comprising these compounds, are useful for the treatment ofcancer and neoplastic disease.

One embodiment provides a compound having the structure of Formula (I),or a pharmaceutically acceptable salt thereof,

wherein,

W is N, C—H, or C—F;

X is hydrogen, halogen, —CN, optionally substituted alkyl, optionallysubstituted alkynyl, optionally substituted carbocyclylalkynyl,optionally substituted aryl, or optionally substituted heteroaryl;

Y is hydrogen, optionally substituted alkyl, optionally substitutedcycloalkyl, or optionally substituted cycloalkylalkyl;

Z is an optionally substituted group chosen from alkyl, carbocyclyl,C-attached heterocyclyl, N-attached heterocyclyl, heterocyclylalkyl,heterocyclylalkenyl, —O-heterocyclyl, —N(R)-heterocyclyl,—O-heterocyclylalkyl, —N(R)-heterocyclylalkyl, —N(R)(C1-C4alkylene)-NR2,—O(C₁-C₄alk-ylene)-NR₂, and R is hydrogen or C₁-C₄alkyl.

One embodiment provides a compound of Formula (I) having the structureof Formula (Ia), or a pharmaceutically acceptable salt thereof,

wherein,

W is N, C—H, or C—F;

X is hydrogen, halogen, —CN, optionally substituted alkynyl, optionallysubstituted carbocyclylalkynyl, optionally substituted aryl, oroptionally substituted heteroaryl;

Y is hydrogen, optionally substituted alkyl, optionally substitutedcycloalkyl, or optionally substituted cycloalkylalkyl; and

Z is an optionally substituted group chosen from N-attachedheterocyclyl, —O— heterocyclylalkyl, —N(H)-heterocyclyl,—N(Me)-heterocyclyl, —N(H)-heterocyclylalkyl, or—N(Me)-heterocyclylalkyl.

One embodiment provides a compound of Formula (I) or (Ia) having thestructure of Formula (Ib), or a pharmaceutically acceptable saltthereof,

wherein,

W is N, C—H, or C—F;

X is hydrogen, halogen, optionally substituted alkynyl, optionallysubstituted carbocyclylalkynyl, optionally substituted aryl, oroptionally substituted heteroaryl;

Y is hydrogen, optionally substituted alkyl, or optionally substitutedcycloalkyl; and

Z is an optionally substituted group chosen from N-heterocyclyl,—O-heterocyclylalkyl, —N(H)-heterocyclylalkyl, or—N(Me)-heterocyclylalkyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein W is C—H. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein W is C—F. Another embodiment providesthe compound of Formula (Ib), or a pharmaceutically acceptable saltthereof, wherein W is N.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein X is hydrogen. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein X is halogen. Another embodimentprovides the compound of Formula (Ib), or a pharmaceutically acceptablesalt thereof, wherein X is optionally substituted alkynyl. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein X is optionally substitutedcarbocyclylalkynyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein X is optionallysubstituted aryl, or optionally substituted heteroaryl. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein X is optionally substituted aryl.Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein X is an optionallysubstituted phenyl. Another embodiment provides the compound of Formula(Ib), or a pharmaceutically acceptable salt thereof, wherein X isoptionally substituted heteroaryl. Another embodiment provides thecompound of Formula (Ib), or a pharmaceutically acceptable salt thereof,wherein X is chosen from an optionally substituted pyridinyl, optionallysubstituted pyrazolyl, or optionally substituted indazolyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Y is hydrogen. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein Y is optionally substituted cycloalkyl.Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Y is optionallysubstituted alkyl. Another embodiment provides the compound of Formula(Ib), or a pharmaceutically acceptable salt thereof, wherein Y is anoptionally substituted C₁-C₃ alkyl. Another embodiment provides thecompound of Formula (Ib), or a pharmaceutically acceptable salt thereof,wherein Y is an optionally substituted C₁ alkyl. Another embodimentprovides the compound of Formula (Ib), or a pharmaceutically acceptablesalt thereof, wherein Y is a methyl group.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —O-heterocyclylalkyl. Another embodiment provides thecompound of Formula (Ib), or a pharmaceutically acceptable salt thereof,wherein Z is an optionally substituted —N(H)-heterocyclylalkyl. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein Z is an optionally substituted—N(Me)-heterocyclylalkyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —O-heterocyclylalkyl and the heterocyclylalkyl group has theformula —R^(c)-heterocyclyl and the R^(c) is an optionally substitutedC₁-C₃ alkylene chain. Another embodiment provides the compound ofFormula (Ib), or a pharmaceutically acceptable salt thereof, wherein Zis an optionally substituted —O-heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁ alkylene chain.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —O-heterocyclylalkyl and the heterocyclylalkyl group has theformula —R^(c)-heterocyclyl and the heterocyclyl is an optionallysubstituted nitrogen-containing 4-, 5-, 6-, or 7-membered heterocyclyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —N(H)-heterocyclylalkyl and the heterocyclylalkyl group hasthe formula —R^(c)-heterocyclyl and the R^(c) is an optionallysubstituted C₁-C₃ alkylene chain. Another embodiment provides thecompound of Formula (Ib), or a pharmaceutically acceptable salt thereof,wherein Z is an optionally substituted —N(H)-heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁ alkylene chain.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —N(H)-heterocyclylalkyl and the heterocyclylalkyl group hasthe formula —R^(c)-heterocyclyl and the heterocyclyl is an optionallysubstituted nitrogen-containing 4-, 5-, 6-, or 7-membered heterocyclyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —N(Me)-heterocyclylalkyl and the heterocyclylalkyl group hasthe formula —R^(c)-heterocyclyl and the R^(c) is an optionallysubstituted C₁-C₃ alkylene chain. Another embodiment provides thecompound of Formula (Ib), or a pharmaceutically acceptable salt thereof,wherein Z is an optionally substituted —N(Me)-heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁ alkylene chain.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted —N(Me)-heterocyclylalkyl and the heterocyclylalkyl group hasthe formula —R^(c)-heterocyclyl and the heterocyclyl is an optionallysubstituted nitrogen-containing 4-, 5-, 6-, or 7-membered heterocyclyl.

Another embodiment provides the compound of Formula (Ib), or apharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted N-heterocyclyl. Another embodiment provides the compound ofFormula (Ib), or a pharmaceutically acceptable salt thereof, wherein Zis a 4-, 5-, 6-, or 7-membered N-heterocyclyl. Another embodimentprovides the compound of Formula (Ib), or a pharmaceutically acceptablesalt thereof, wherein Z is a 6-membered N-heterocyclyl. Anotherembodiment provides the compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, wherein Z is an optionally substitutedpiperidine. Another embodiment provides the compound of Formula (Ib), ora pharmaceutically acceptable salt thereof, wherein Z is an optionallysubstituted 4-aminopiperidine.

In some embodiments, the substituted heterocyclic derivative compounddescribed in Formula (I), (Ia), or (Ib) has a structure provided inTable 1.

TABLE 1 Chemical Synthesis Example Structure Name 1

4-(2-(4-aminopiperidin-1-yl)- 1-methyl-6-oxo-5-p-tolyl-1,6-dihydropyrimidin-4-yl)-benzonitrile 2

4-[2-(4-amino-piperidin-1-yl)-5- (4-methoxyphenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- benzonitrile 3

4-[2-(4-amino-piperidin-1-yl)-5- (6-methoxypyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- benzonitrile 4

4-[2-(4-amino-piperidin-1-yl)-1- methyl-5-(6-methylpyridin-3-yl)-6-oxo-1,6-dihydro-pyrimidin-4- yl]-benzonitrile 5

4-[2-(4-amino-piperidin-1-yl)-5-(4- methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-4-yl]- benzonitrile 6

4-[2-(4-amino-piperidin-1-yl)-5- (4-methoxyphenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluorobenzonitrile 7

4-[2-(4-amino-piperidin-1-yl)-5- (3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 8

4-[2-(4-amino-piperidin-1-yl)-5- (6-methoxypyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 9

4-[2-(4-amino-piperidin-1-yl)-5- (6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4- yl]-2-fluoro-benzonitrile 10

4-[2-(4-amino-piperidin-1-yl)-5-(6- ethyl-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- benzonitrile 11

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-2-(4-methylamino-piperidin-1-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 12

2-fluoro-4-[5-(3-fluoro-4-methoxy- phenyl)-1-methyl-2-(4-methyl-amino-piperidin-1-yl)-6-oxo-1,6- dihydro-pyrimidin-4-yl]- benzonitrile13

4-[2-(4-amino-piperidin-1-yl)-1- ethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro- benzonitrile 14

4-[2-(4-amino-piperidin-1-yl)-5- cyclopentylethynyl-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 15

[2-(4-amino-piperidin-1-yl)-4-(4- cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H- pyrimidin-1-yl]-acetic acid 16

2-[2-(4-amino-piperidin-1-yl)-4- (4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H- pyrimidin-1-yl]-acetamide 17

4-[2-(4-amino-piperidin-1-yl)-1- (3-hydroxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro- benzonitrile 18

4-[2-(4-amino-piperidin-1-yl)-5- benzofuran-5-yl-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2- fluoro-benzonitrile 19

2-(4-amino-piperidin-1-yl)-4-(4- cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidine-5- carbonitrile 20

4-[2-(4-aminopiperidin-1-yl)-5- chloro-1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 21

2-fluoro-4-[1-methyl-2- (4-methylamino-piperidin-1-yl)-5-(6-methyl-pyridin-3-yl)-6-oxo- 1,6-dihydro-pyrimidin-4-yl]- benzonitrile22

4-[2-(2,8-diaza-spiro[4.5]dec-8-yl)- 5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2- fluorobenzonitrile 23

4-{2-(4-aminopiperidyl)-1-methyl- 6-oxo-5-[6-(trifluoromethyl) (3-pyridyl)]hydropyrimidin-4-yl}- 2-fluorobenzenecarbonitrile 24

4-[2-(4-aminopiperidyl)-1-methyl- 5-(2-methyl(2H-indazol-5-yl))-6-oxohydropyrimidin-4-yl] benzenecarbonitrile 25

4-[2-((3R)-3-aminopiperidyl)-5- (3-fluoro-4-methoxyphenyl)-1-methyl-6-oxohydropyrimidin-4- yl]-2-fluorobenzenecarbonitrile 26

4-[2-(4-aminopiperidyl)-5-(5- fluoro-6-methoxy(3-5,6-dihydropyridyl))-1-methyl-6- oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 27

4-[2-((3R)-3-aminopyrrolidinyl)- 5-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxohydropyrimidin- 4-yl]-2-fluorobenzenecarbonitrile 28

4-[2-((3S)-3-amino-piperidin-1- yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 29

4-[2-((3S)-3-amino-pyrrolidin-1-yl)- 5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 30

4-[2-((3R)-3-aminopiperidyl)-5- (4-methoxyphenyl)-1-methyl-6-oxohydro-pyrimidin-4-yl]-2- fluorobenzenecarbonitrile 31

4-[2-((3S)-3-amino-piperidin-1- yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 32

4-[2-(4-amino-4-methyl-piperidin- 1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 33

4-[2-(4-aminopiperidyl)-1-methyl- 5-(1-methyl(1H-indazol-5-yl))-6-oxohydropyrimidin-4-yl] benzenecarbonitrile 34

4-{2-(4-amino-piperidin-1-yl)-1- methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-1H-pyrazol-4-yl]-1,6- dihydro-pyrimidin-4-yl}-2-fluoro-benzonitrile 35

4-[2-(4-amino-piperidin-1-yl)-1- methyl-5-(1-methyl-1H-indazol-5-yl)-6-oxo-1,6-dihydro-pyrimidin- 4-yl]-2-fluoro-benzonitrile 36

4-{2-(4-amino-piperidin-1-yl)-1- methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-1H-pyrazol-4-yl]- 1,6-dihydro-pyrimidin-4-yl}-benzonitrile 37

4-[2-(4-aminopiperidyl)-1-methyl- 5-(2-methyl(2H-indazol-5-yl))-6-oxohydropyrimidin-4-yl]- 2-fluorobenzenecarbonitrile 38

4-[2-(4-aminopiperidyl)-5- (3,5-difluoro-4-methoxyphenyl)-1-methyl-6-oxohydropyrimidin- 4-yl]benzenecarbonitrile 39

4-[2-(4-aminopiperidyl)-6-(4- cyano-3-fluorophenyl)-3-methyl-4-oxo-3-hydropyrimidin-5-yl] benzoic acid 40

{4-[2-(4-aminopiperidyl)-6- (4-cyanophenyl)-3-methyl-4-oxo (3-hydropyrimidin-5-yl)]-2- fluorophenyl}-N- methylcarboxamide 41

4-[2-(4-aminopiperidyl)-6- (4-cyanophenyl)-3-methyl-4- oxo(3-hydropyrimidin-5-yl)]-2- fluorobenzamide 42

4-[2-(4-amino-piperidin-1-yl)-1- methyl-6-oxo-5-(1-oxo-2,3-dihydro-1H-isoindol-5-yl)-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 43

3-[2-(4-amino-piperidin-1-yl)-4- (4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-5-yl]-benzoic acid 44

4-{5-(3-fluoro-4-methoxy-phenyl)- 1-methyl-6-oxo-2-[(3S)-(pyrrolidin-3-ylmethyl)-amino]-1,6-dihydro- pyrimidin-4-yl}-benzonitrile 45

4-{5-(3-fluoro-4-methoxy-phenyl)- 1-methyl-6-oxo-2-[(3R)-(pyrrolidin-3-ylmethyl)-amino]- 1,6-dihydro-pyrimidin-4-yl}-benzonitrile 46

4-[2-[1,4]diazepan-1-yl-5-(3-fluoro- 4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 47

2-fluoro-4-[5-(3-fluoro- 4-methoxy-phenyl)-1-methyl-6-oxo-2-piperazin-1-yl-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 48

4-[5-(3-fluoro-4-methoxy-phenyl)- 1-methyl-6-oxo-2-(piperidin-4-ylamino)-1,6-dihydro-pyrimidin-4- yl]-benzonitrile 49

4-[2-(4-amino-piperidin-1-yl)- 2′-dimethylamino-1-methyl-6-oxo-1,6-dihydro-[5,5′]bipyrimidinyl-4- yl]-2-fluoro-benzonitrile 50

5-[2-(4-amino-piperidin-1-yl)-4- (4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-5-yl]- pyridine-2-carboxylic acidmethylamide 51

2-fluoro-4-{5-(4-methoxy-phenyl)- 1-methyl-6-oxo-2-[(3S)-(pyrrolidin-3-ylmethyl)-amino]-1,6-dihydro- pyrimidin-4-yl}-benzonitrile 52

2-fluoro-4-{5-(4-methoxy-phenyl)- 1-methyl-6-oxo-2-[(3R)-(pyrrolidin-3-ylmethyl)-amino]-1,6-dihydro- pyrimidin-4-yl}-benzonitrile 53

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-6-oxo-2-(piperidin-4-ylamino)-1,6-dihydro-pyrimidin-4- yl]-benzonitrile 54

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-2-(methyl-(3S)-pyrrolidin-3-ylmethyl-amino)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 55

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-2-(methyl-piperidin-4-yl-amino)-6-oxo-1,6-dihydro-pyrimidin- 4-yl]-benzonitrile 56

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-2-(methyl-pyrrolidin-3-ylmethyl-amino)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 57

4-[2-(4-amino-piperidin-1-yl)-5- (6-dimethylamino-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 58

2-fluoro-4-[5-(6-methoxy-pyridin-3- yl)-1-methyl-2-(4-methylamino-piperidin-1-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 59

4-[2-(4-amino-piperidin-1-yl)-5- (4-dimethylamino-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 60

4-[2-(4-amino-piperidin-1-yl)-1- methyl-6-oxo-5-(6-pyrrolidin-1-yl-pyridin-3-yl)-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 61

4-[2-[1,4]diazepan-1-yl-5-(6- methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4- yl]-2-fluoro benzonitrile 62

4-[2-[1,4]diazepan-1-yl-5-(6- methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4- yl]-2-fluoro-benzonitrile 63

4-[2-[1,4]diazepan-1-yl-5- (6-dimethylamino-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile 64

4-[2-(3-amino-azetidin-1-yl)-5- (4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 65

2-fluoro-4-[1-methyl-2- (4-methylamino-piperidin-1-yl)-5-(2-methyl-2H-indazol-5-yl)-6-oxo- 1,6-dihydro-pyrimidin-4-yl]-benzonitrile 66

4-[2-[1,4]diazepan-1-yl-1-methyl- 5-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 67

4-[2-[1,4]diazepan-1-yl-5- (6-dimethylamino-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 68

4-[2-(4-amino-piperidin-1-yl)-1- methyl-5-(6-morpholin-4-yl-pyridin-3-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile69

4-[2-(3-aminomethyl-azetidin-1-yl)- 5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 70

2-fluoro-4-[5-(4-methoxy-phenyl)- 1-methyl-2-(3-methylaminomethyl-azetidin-1-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-benzonitrile 71

4-[2-(4-dimethylamino-piperidin-1- yl)-1-methyl-5-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile72

4-[2-(4-dimethylamino-piperidin-1- yl)-1-methyl-5-(1-methyl-1H-indazol-5-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile73

4-[2-(4-amino-piperidin-1-yl)-5- (1H-indol-5-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 74

4-[2-(4-amino-piperidin-1-yl)-1- methyl-5-(1-methyl-1H-indol-5-yl)-6-oxo-1,6-dihydro-pyrimidin- 4-yl]-2-fluoro-benzonitrile 75

4-[2-(4-amino-piperidin-1-yl)-5- (1H-indol-6-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl)-2- fluoro-benzonitrile 76

4-[2-(4-amino-piperidin-1-yl)-1- methyl-5-(1-methyl-1H-indol-6-yl)-6-oxo-1,6-dihydro-pyrimidin- 4-yl]-2-fluoro-benzonitrile 77

4-[2-(4-amino-piperidin-1-yl)-5- (1H-indazol-6-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 78

4-[2-((4R,3S)-4-amino-3-fluoro- piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 79

4-[2-((4S,3R)-4-amino-3-fluoro- piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 80

4-[2-(4-dimethylamino-piperidin-1- yl)-1-methyl-5-(2-methyl-2H-indazol-6-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro- benzonitrile81

4-[2′-dimethylamino-2-(4- dimethylamino-piperidin-1-yl)-1-methyl-6-oxo-1,6-dihydro- [5,5′]bipyrimidinyl-4-yl]-2-fluoro-benzonitrile 82

4-[2-(4-dimethylamino-piperidin-1- yl)-1-methyl-5-(6-methyl-pyridin-3-yl)-6-oxo-1,6-dihydro-pyrimidin- 4-yl]-2-fluoro-benzonitrile 83

4-[5-(6-dimethylamino-pyridin-3- yl)-1-methyl-2-(4-methylamino-piperidin-1-yl)-6-oxo-1,6-dihydro- pyrimidin-4-yl]-2-fluoro-benzonitrile 84

4-[2-(4-dimethylamino-piperidin-1- yl)-5-(2H-indazol-6-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]- 2-fluoro-benzonitrile 85

4-[2-(4-amino-piperidin-1-yl)-5-(3- fluoro-4-methoxy-phenyl)-1-deuteratedmethyl-6-oxo-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 86

4-[2-(4-amino-piperidin-1-yl)-5- (3-fluoro-4-deuteratedmethoxy-phenyl)-1-methyl-6-oxo-1,6- dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 87

2-fluoro-4-[1-methyl-2- [4-(methylamino)piperidin-1-yl]-5-(1-methylindazol-5-yl)-6- oxopyrimidin-4-yl] benzonitrile 88

4-[2-(4-aminopiperidin-1-yl)-5- (1H-indazol-5-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 89

4-[5-(4-aminophenyl)-2-(4- aminopiperidin-1-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 90

4-[2-(4-aminopiperidin-1-yl)-1- methyl-5-[4-(methylamino)phenyl]-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 91

4-[2-(4-aminopiperidin-1-yl)-5-[3- fluoro-4-(methylamino)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 92

4-[2-[4-(dimethylamino)piperidin- 1-yl]-5-(6-methoxypyridin-3-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 93

4-[2-(4-aminopiperidin-1-yl)-5-(6- ethoxy-5-fluoropyridin-3-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 94

4-[2-(4-aminopiperidin-1-yl)-5-(6- ethoxypyridin-3-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 95

4-[2-(4-aminopiperidin-1-yl)-5-(4- ethoxyphenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 96

4-[2-(4-aminopiperidin-1-yl)-5-[4-(2- hydroxyethoxy)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 97

4-[2-(4-aminopiperidin-1-yl)-5-[4-(2- hydroxyethoxy)phenyl]-1-methyl-6-oxopyrimidin-4-yl]benzonitrile 98

4-[2-(4-aminopiperidin-1-yl)-5-[4-(2- methoxyethoxy)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 99

4-[2-(4-aminopiperidin-1-yl)-5-[4-(2- hydroxyethyl)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 100

4-[2-(4-aminopiperidin-1-yl)-5-[4- (hydroxymethyl)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 101

4-[2-(4-aminopiperidin-1-yl)-5-(4- fluorophenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 102

4-[2-(4-aminopiperidin-1-yl)-5-(3- fluorophenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 103

4-[2-(4-aminopiperidin-1-yl)-5-(3,5- difluorophenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 104

4-[2-(4-aminopiperidin-1-yl)-5-(3,4- difluorophenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 105

4-[2-(4-aminopiperidin-1-yl)-1- methyl-5-(4-methylsulfonylphenyl)-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 106

4-[2-(4-aminopiperidin-1-yl)-5-(4- chlorophenyl)-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 107

4-[2-(4-aminopiperidin-1-yl)-5-[4- (methoxymethyl)phenyl]-1-methyl-6-oxopyrimidin-4-yl]-2- fluorobenzonitrile 108

4-[2-(4-aminopiperidin-1-yl)-1- methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 109

4-[2-(4-amino-piperidin-1-yl)-1- cyclopropylmethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2- fluoro-benzonitrile 110

4-[2-(4-amino-piperidin-1-yl)-1- cyclopropylmethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2- fluoro-benzonitrile 111

2-(4-amino-piperidin-1-yl)-6-(4- chloro-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-3-methyl-3H- pyrimidin-4-one 112

2-(4-amino-piperidin-1-yl)-6-(4- hydroxy-phenyl)-3-methyl-5-(1-methyl-1H-indol-5-yl)-3H- pyrimidin-4-one 113

2-(4-amino-piperidin-1-yl)-6-(4- fluoro-phenyl)-3-methyl-5-(1-methyl-1H-indol-5-yl)-3H- pyrimidin-4-one 114

2-(4-amino-piperidin-1-yl)-3- methyl-5-(1-methyl-1H-indol-5-yl)-6-phenyl-3H-pyrimidin-4- one 115

2-(4-amino-piperidin-1-yl)-5-(3- fluoro-4-methoxy-phenyl)-3-methyl-6-pyridin-4-yl-3H- pyrimidin-4-one 116

2-(4-amino-piperidin-1-yl)-3- methyl-5-(1-methyl-1H-indol-5-yl)-6-pyridin-4-yl-3H- pyrimidin-4-one 117

2-(4-amino-piperidin-1-yl)-6-(4- methoxy-phenyl)-3-methyl-5-(1-methyl-1H-indol-5-yl)-3H- pyrimidin-4-one 118

3-[2-(4-aminopiperidin-1-yl)-5-(3- fluoro-4-methoxyphenyl)-1-methyl-6-oxopyrimidin-4-yl]benzonitrile 119

2-[2-(4-aminopiperidin-1-yl)-5-(3- fluoro-4-methoxyphenyl)-1-methyl-6-oxopyrimidin-4-yl]benzonitrile 120

2-(4-amino-piperidin-1-yl)-5-(3- fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro- pyrimidine-4-carbonitrile 121

2-(4-amino-piperidin-1-yl)-4-(4- cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidine- 5-carbonitrile 122

4-[2-(4-aminopiperidin-1-yl)-5-(4- methoxyphenyl)-6-oxo-1H-pyrimidin-4-yl]-2- fluorobenzonitrile

In some embodiments, the substituted heterocyclic derivative compounddescribed herein has the structure provided in Table 2.

TABLE 2

Preparation of the Substituted Heterocyclic Derivative Compounds

The compounds used in the reactions described herein are made accordingto organic synthesis techniques known to those skilled in this art,starting from commercially available chemicals and/or from compoundsdescribed in the chemical literature. “Commercially available chemicals”are obtained from standard commercial sources including Acros Organics(Pittsburgh, Pa.), Aldrich Chemical (Milwaukee, Wis., including SigmaChemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), AvocadoResearch (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet(Cornwall, U.K.), Chemservice Inc. (West Chester, Pa.), CrescentChemical Co. (Hauppauge, N.Y.), Eastman Organic Chemicals, Eastman KodakCompany (Rochester, N.Y.), Fisher Scientific Co. (Pittsburgh, Pa.),Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan,Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.), Key Organics(Cornwall, U.K.), Lancaster Synthesis (Windham, N.H.), MaybridgeChemical Co. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, Utah),Pfaltz & Bauer, Inc. (Waterbury, Conn.), Polyorganix (Houston, Tex.),Pierce Chemical Co. (Rockford, Ill.), Riedel de Haen AG (Hanover,Germany), Spectrum Quality Product, Inc. (New Brunswick, N.J.), TCIAmerica (Portland, Oreg.), Trans World Chemicals, Inc. (Rockville, Md.),and Wako Chemicals USA, Inc. (Richmond, Va.).

Suitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, SYNTHETIC ORGANIC CHEMISTRY, John Wiley & Sons, Inc., NewYork; S. R. Sandler et al., ORGANIC FUNCTIONAL GROUP PREPARATIONS, 2ndEd., Academic Press, New York, 1983; H. O. House, MODERN SYNTHETICREACTIONS, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L.Gilchrist, HETEROCYCLIC CHEMISTRY, 2nd Ed., John Wiley & Sons, New York,1992; J. March, ADVANCED ORGANIC CHEMISTRY: REACTIONS, MECHANISMS &STRUCTURE, 4th Ed., Wiley-Interscience, New York, 1992. Additionalsuitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, Fuhrhop, J. and Penzlin G., ORGANIC SYNTHESIS: CONCEPTS,METHODS, STARTING MATERIALS, SECOND, REVISED & ENLARGED EDITION (1994)John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V., ORGANICCHEMISTRY, AN INTERMEDIATE TEXT (1996) Oxford University Press, ISBN0-19-509618-5; Larock, R. C. COMPREHENSIVE ORGANIC TRANSFORMATIONS: AGUIDE TO FUNCTIONAL GROUP PREPARATION, 2nd Ed. (1999) Wiley-VCH, ISBN:0-471-19031-4; March, J. ADVANCED ORGANIC CHEMISTRY: REACTIONS,MECHANISMS, & STRUCTURE, 4th Ed. (1992) John Wiley & Sons, ISBN:0-471-60180-2; Otera, J. (ed.), MODERN CARBONYL CHEMISTRY, (2000)Wiley-VCH, ISBN: 3-527-29871-1; Patai, S., PATAI'S 1992 GUIDE TO THECHEMISTRY OF FUNCTIONAL GROUPS, (1992) Interscience ISBN: 0-471-93022-9;Solomons, T. W. G., ORGANIC CHEMISTRY, 7th Ed. (2000) John Wiley & Sons,ISBN: 0-471-19095-0; Stowell, J. C., INTERMEDIATE Organic Chemistry, 2ndEd. (1993) Wiley-Interscience, ISBN: 0-471-57456-2; INDUSTRIAL ORGANICCHEMICALS: STARTING MATERIALS & INTERMEDIATES: AN ULLMANN'SENCYCLOPEDIA, (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8volumes; ORGANIC REACTIONS (1942-2000) John Wiley & Sons, in over 55volumes; and CHEMISTRY OF FUNCTIONAL GROUPS, John Wiley & Sons, in 73volumes.

Specific and analogous reactants are optionally identified through theindices of known chemicals prepared by the Chemical Abstract Service ofthe American Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (contact theAmerican Chemical Society, Washington, D.C. for more details). Chemicalsthat are known but not commercially available in catalogs are optionallyprepared by custom chemical synthesis houses, where many of the standardchemical supply houses (e.g., those listed above) provide customsynthesis services. A reference for the preparation and selection ofpharmaceutical salts of the substituted heterocyclic derivativecompounds described herein is P. H. Stahl & C. G. Wermuth, HANDBOOK OFPHARMACEUTICAL SALTS, Verlag Helvetica Chimica Acta, Zurich, 2002.

The substituted heterocyclic derivative compounds are prepared by thegeneral synthetic route described below in Scheme 1.

Referring to Scheme 1, compound A is selectively hydrolyzed to givecompound B. Compound C is obtained from N-alkylation of compound B witha variety of alkyl halides R₁—X. Selective displacement of trichloridecompound C is carried out with a variety of amines HN(R₂)(R₂′) underbasic conditions to form compound D. Compound E is prepared fromcompound D under palladium-mediated cross coupling conditions withboronic acids, e.g. R₃—B(OH)₂, or boronic esters. Compound F is preparedfrom compound E under palladium-mediated cross coupling conditions withboronic acids, e.g., R₃—B(OH)₂, or boronic esters.

Pharmaceutical Compositions of the Substituted Heterocyclic DerivativeCompounds

In certain embodiments, the substituted heterocyclic derivative compoundas described herein is administered as a pure chemical. In otherembodiments, the substituted heterocyclic derivative compound describedherein is combined with a pharmaceutically suitable or acceptablecarrier (also referred to herein as a pharmaceutically suitable (oracceptable) excipient, physiologically suitable (or acceptable)excipient, or physiologically suitable (or acceptable) carrier) selectedon the basis of a chosen route of administration and standardpharmaceutical practice as described, for example, in REMINGTON: THESCIENCE & PRACTICE OF PHARMACY (Gennaro, 21^(st) Ed. Mack Pub. Co.,Easton, Pa. (2005)).

Provided herein is a pharmaceutical composition comprising at least onesubstituted heterocyclic derivative compound, or a stereoisomer,pharmaceutically acceptable salt, hydrate, solvate, or N-oxide thereof,together with one or more pharmaceutically acceptable carriers. Thecarrier(s) (or excipient(s)) is acceptable or suitable if the carrier iscompatible with the other ingredients of the composition and notdeleterious to the recipient (i.e., the subject) of the composition.

One embodiment provides a pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient. One embodiment provides apharmaceutical composition comprising a compound of Formula (Ia), or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient. One embodiment provides a pharmaceuticalcomposition comprising a compound of Formula (Ib), or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable excipient.

In certain embodiments, the substituted heterocyclic derivative compoundas described by Formula (I) is substantially pure, in that it containsless than about 5%, or less than about 1%, or less than about 0.1%, ofother organic small molecules, such as unreacted intermediates orsynthesis by-products that are created, for example, in one or more ofthe steps of a synthesis method.

Suitable oral dosage forms include, for example, tablets, pills,sachets, or capsules of hard or soft gelatin, methylcellulose or ofanother suitable material easily dissolved in the digestive tract. Insome embodiments, suitable nontoxic solid carriers are used whichinclude, for example, pharmaceutical grades of mannitol, lactose,starch, magnesium stearate, sodium saccharin, talcum, cellulose,glucose, sucrose, magnesium carbonate, and the like. (See, e.g.,REMINGTON: THE SCIENCE & PRACTICE OF PHARMACY (Gennaro, 21^(st) Ed. MackPub. Co., Easton, Pa. (2005)).

The dose of the composition comprising at least one substitutedheterocyclic derivative compound as described herein differ, dependingupon the patient's (e.g., human) condition, that is, stage of thedisease, general health status, age, and other factors.

Pharmaceutical compositions are administered in a manner appropriate tothe disease to be treated (or prevented). An appropriate dose and asuitable duration and frequency of administration will be determined bysuch factors as the condition of the patient, the type and severity ofthe patient's disease, the particular form of the active ingredient, andthe method of administration. In general, an appropriate dose andtreatment regimen provides the composition(s) in an amount sufficient toprovide therapeutic and/or prophylactic benefit (e.g., an improvedclinical outcome, such as more frequent complete or partial remissions,or longer disease-free and/or overall survival, or a lessening ofsymptom severity. Optimal doses are generally determined usingexperimental models and/or clinical trials. The optimal dose dependsupon the body mass, weight, or blood volume of the patient.

Oral doses typically range from about 1.0 mg to about 1000 mg, one tofour times, or more, per day.

Use of the Substituted Heterocyclic Derivative Compounds

Epigenetics is the study of heritable changes in gene expression causedby mechanisms other than the underlying DNA sequence. Molecularmechanisms that play a role in epigenetic regulation include DNAmethylation and chromatin/histone modifications.

The genomes of eukaryotic organisms are highly organized within thenucleus of the cell. Tremendous compaction is required to package the 3billion nucleotides of the human genome into the nucleus of a cell.Chromatin is the complex of DNA and protein that makes up chromosomes.Histones are the major protein component of chromatin, acting as spoolsaround which DNA winds. Changes in chromatin structure are affected bycovalent modifications of histone proteins and by non-histone bindingproteins. Several classes of enzymes are known which modify histones atvarious sites.

There are a total of six classes of histones (HI, H2A, H2B, H3, H4, andH5) organized into two groups: core histones (H2A, H2B, H3, and H4) andlinker histones (HI and H5). The basic unit of chromatin is thenucleosome, which consists of about 147 base pairs of DNA wrapped aroundthe core histone octamer, consisting of two copies each of the corehistones H2A, H2B, H3, and H4.

Basic nucleosome units are then further organized and condensed by theaggregation and folding of nucleosomes to form a highly condensedchromatin structure. A range of different states of condensation arepossible, and the tightness of chromatin structure varies during thecell cycle, being most compact during the process of cell division.

Chromatin structure plays a critical role in regulating genetranscription, which cannot occur efficiently from highly condensedchromatin. The chromatin structure is controlled by a series of posttranslational modifications to histone proteins, notably histones H3 andH4, and most commonly within the histone tails which extend beyond thecore nucleosome structure. These modifications are acetylation,methylation, phosphorylation, ribosylation sumoylation, ubiquitination,citrullination, deimination, and biotinylation. The core of histones H2Aand H3 can also be modified. Histone modifications are integral todiverse biological processes such as gene regulation, DNA repair, andchromosome condensation.

Histone methylation is one of the most important chromatin marks; theseplay important roles in transcriptional regulation, DNA-damage response,heterochromatin formation and maintenance, and X-chromosomeinactivation. A recent discovery also revealed that histone methylationaffects the splicing outcome of pre-mRNA by influencing the recruitmentof splicing regulators. Histone methylation includes mono-, di-, andtri-methylation of lysines, and mono-, symmetric di-, and asymmetricdi-methylation of arginines. These modifications can be either anactivating or repressing mark, depending on the site and degree ofmethylation.

Histone Demethylases

A “demethylase” or “protein demethylase,” as referred to herein, refersto an enzyme that removes at least one methyl group from polypeptide.Demethylases comprise a JmjC domain, and can be a methyl-lysine ormethyl-arginine demethylase. Some demethylases act on histones, e.g.,act as a histone H3 or H4 demethylase. For example, an H3 demethylasemay demethylate one or more of H3K4, H3K9, H3K27, H3K36 and/or H3K79.Alternately, an H4 demethylase may demethylate histone H4K20.Demethylases are known which can demethylate either a mono-, di- and/ora tri-methylated substrate. Further, histone demethylases can act on amethylated core histone substrate, a mononucleosome substrate, adinucleosome substrate and/or an oligonucleosome substrate, peptidesubstrate and/or chromatin (e.g., in a cell-based assay).

The first lysine demethylase discovered was lysine specific demethylase1 (LSD1/KDM1), which demethylates both mono- and di-methylated H3K4 orH3K9, using flavin as a cofactor. A second class of Jumonji C (JmjC)domain containing histone demthylases were predicted, and confirmed whena H3K36 demethylase was found used a formaldehyde release assay, whichwas named JmjC domain containing histone demethylase 1 (JHDM1/KDM2A).

More JmjC domain-containing proteins were subsequently identified andthey can be phylogenetically clustered into seven subfamilies: JHDM1,JHDM2, JHDM3, JMJD2, JARID, PHF2/PHF8, UTX/UTY, and JmjC domain only.

LSD-1

Lysine-specific demethylase 1 (LSD1) is a histone lysine demethylasethat specifically demethylates monomethylated and dimethylated histoneH3 at K4 and also demethylates dimethylated histone H3 at K9. Althoughthe main target of LSD1 appears to be mono- and di-methylated histonelysines, specifically H3K4 and H3K9, there is evidence in the literaturethat LSD 1 can demethylate methylated lysines on non-histone proteinslike p53, E2F1, Dnmtl and STAT3.

LSD 1 has a fair degree of structural similarity and amino acididentity/homology to polyamine oxidases and monoamine oxidases, all ofwhich (i.e., MAO-A, MAO-B and LSD1) are flavin dependent amine oxidaseswhich catalyze the oxidation of nitrogen-hydrogen bonds and/ornitrogen-carbon bonds. LSD1 also includes an N-terminal SWRIM domain.There are two transcript variants of LSD1 produced by alternativesplicing.

In some embodiments, the compounds disclosed herein are capable ofinhibiting LSD1 activity in a biological sample by contacting thebiological sample with a substituted heterocyclic compound as disclosedherein. In some embodiments, a substituted heterocyclic compound asdisclosed herein is capable of modulating the level of histone-4lysine-3 methylation in the biological sample. In some embodiments, asubstituted heterocyclic compound as disclosed herein is capable ofmodulating histone-3 lysine-9 methylation levels in the biologicalsample.

The substituted heterocyclic compounds disclosed herein lack significantMAO-A or MAO-B inhibitory activity. In some embodiments, a substitutedheterocyclic compound as disclosed herein inhibits LSD1 inhibitoryactivity to a greater extent than MAO-A and/or MAO-B inhibitoryactivity.

One embodiment provides a method of regulating gene transcription in acell comprising inhibiting lysine-specific demethylase 1 activity byexposing the lysine-specific demethylase 1 enzyme to a compound ofFormula (I). One embodiment provides a method of regulating genetranscription in a cell comprising inhibiting lysine-specificdemethylase 1 activity by exposing the lysine-specific demethylase 1enzyme to a compound of Formula (Ia). One embodiment provides a methodof regulating gene transcription in a cell comprising inhibitinglysine-specific demethylase 1 activity by exposing the lysine-specificdemethylase 1 enzyme to a compound of Formula (Ib).

Methods of Treatment

Disclosed herein are methods of modulating demethylation in a cell or ina subject, either generally or with respect to one or more specifictarget genes. Demethylation is modulated to control a variety ofcellular functions, including without limitation: differentiation;proliferation; apoptosis; tumorigenesis, leukemogenesis or otheroncogenic transformation events; hair loss; or sexual differentiation.

One embodiment provides a method of treating cancer in a patient in needthereof, comprising administering to the patient a compound of Formula(I), or a pharmaceutically acceptable salt thereof. One embodimentprovides a method of treating cancer in a patient in need thereof,comprising administering to the patient a compound of Formula (Ia), or apharmaceutically acceptable salt thereof. One embodiment provides amethod of treating cancer in a patient in need thereof, comprisingadministering to the patient a compound of Formula (Ib), or apharmaceutically acceptable salt thereof.

In a further embodiment is the method for treating cancer in a subjectwherein the cancer is selected from prostate cancer, breast cancer,bladder cancer, lung cancer or melanoma. In a further embodiment is themethod for treating cancer in a subject wherein the cancer is selectedfrom acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL),small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC),neuroblastoma, small round blue cell tumors, or glioblastoma.

Other embodiments and uses will be apparent to one skilled in the art inlight of the present disclosures. The following examples are providedmerely as illustrative of various embodiments and shall not be construedto limit the invention in any way.

EXAMPLES I. Chemical Synthesis

Unless otherwise noted, reagents and solvents were used as received fromcommercial suppliers. Anhydrous solvents and oven-dried glassware wereused for synthetic transformations sensitive to moisture and/or oxygen.Yields were not optimized. Reaction times are approximate and were notoptimized. Column chromatography and thin layer chromatography (TLC)were performed on silica gel unless otherwise noted. Spectra are givenin ppm (δ) and coupling constants, J are reported in Hertz. For protonspectra the solvent peak was used as the reference peak.

Preparation 1A: 2,5,6-trichloropyrimidin-4-ol

To a solution of 2,4,5,6-tetrachloropyrimidine (5 g, 22.9 mmol) in THF(50 mL) was added 1N NaOH (31 mL, 31.2 mmol) dropwise, and the mixturewas stirred overnight at RT. The solution was acidified with 1N HCl andextracted with DCM (3×). The organics were combined, dried, andconcentrated in vacuo. The solids were slurried in Et₂O for 30 min atRT, filtered, washed with Et₂O, and dried to give 3.0 g (66%) of thetitle compound. [M+H] Calc'd for C₄HCl₃N₂O, 201; Found, 201.

Preparation 1B: 2,5,6-trichloro-3-methyl-3-hydropyrimidin-4-one

To a mixture of 2,5,6-trichloropyrimidin-4-ol (1 g, 5.0 mmol) and K₂CO₃(759 mg, 5.5 mmol) in THF (50 mL) at 0° C. was added iodomethane (714mg, 5.0 mmol) dropwise, and the reaction was stirred at RT overnight.The reaction mixture was diluted with ethyl acetate (EA). The organicphase was washed with brine, dried and concentrated in vacuo. Theresidue was purified by silica gel chromatography (10:1, PE:EA) to give760 mg (71%) of the title compound. ¹H NMR (400 MHz, CDCl₃): δ 3.74 (s,3H). [M+H] Calc'd for C₅H₃C₁₃N₂O, 213; Found, 213.

Preparation 1C:N-[1-(5,6-dichloro-3-methyl-4-oxo(3-hydropyrimidin-2-yl))(4-piperidyl)](tert-butoxy)carboxamide

A solution of 2,5,6-trichloro-3-methyl-3-hydropyrimidin-4-one (426 mg,2.0 mmol), DIEA (536 mg, 4.0 mmol) and tert-butylpiperidin-4-ylcarbamate (400 mg, 2 mmol) in DMF (10 mL) was heated at120° C. for 1 h. The solvent was removed in vacuo and the residue waspurified by silica gel chromatography (1:1, PE:EA) to give 550 mg (73%)of the title compound. ¹H NMR (400 MHz, CDCl₃): δ 1.45 (s, 9H),1.50-1.58 (m, 2H), 2.06-2.10 (m, 2H), 2.98-3.05 (m, 2H), 3.48 (s, 3H),3.53-3.56 (m, 2H), 3.70 (s, 1H), 4.52 (s, 1H). [M+H] Calc'd forC₁₅H₂₂Cl₂N₄O₃, 213; Found, 213.

Preparation 1D: tert-butyl1-(5-chloro-4-(4-cyanophenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate

A mixture ofN-[1-(5,6-dichloro-3-methyl-4-oxo(3-hydropyrimidin-2-yl))(4-piperidyl)](tert-butoxy)carboxamide(500 mg, 1.3 mmol), 4-cyanophenylboronic acid (195 mg, 1.3 mmol),[1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (81 mg,0.13 mmol) and K₂CO₃ (359 mg, 2.6 mmol) in DMF (10 mL) was flushed withnitrogen and stirred at 85° C. for 2 h. Water was added, and the mixturewas extracted with EA (3×). The organics were combined, washed withwater, washed with brine, dried and concentrated in vacuo. The residuewas purified purified by silica chromatography (1:1, EA:PE) to give 250mg (40%) of the title compound.

¹H NMR (400 MHz, CDCl₃): δ 1.45 (s, 9H), 1.54-1.61 (m, 2H), 2.05-2.10(m, 2H), 2.99-3.05 (m, 2H), 3.48-3.56 (s, 5H), 3.70 (s, 1H), 4.56 (s,1H), 7.73 (d, J=8.0 Hz, 2H), 7.93 (d, J=8.0 Hz, 2H). [M+H] Calc'd forC₂₂H₂₆ClN₅O₃, 444; Found, 444.

Preparation 1E: tert-butyl1-(4-(4-cyanophenyl)-1-methyl-6-oxo-5-p-tolyl-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate

A mixture of tert-butyl1-(5-chloro-4-(4-cyanophenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate(200 mg, 0.45 mmol), p-tolylboronic acid (123 mg, 0.90 mmol),[1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (28 mg,0.045 mol) and K₂CO₃ (124 mg, 0.90 mmol) in DMF (10 mL) was flushed withnitrogen and stirred at 85° C. for 2 h. Water was added, and the mixturewas extracted with EA (3×). The organics were combined, washed withwater, washed with brine, dried and concentrated in vacuo. The residuewas purified by silica chromatography (1:1, EA:PE) to give 50 mg (22%)of the title compound. [M+H] Calc'd for C₂₉H₃₃N₅O₃, 500; Found, 500.

Example 1:4-(2-(4-aminopiperidin-1-yl)-1-methyl-6-oxo-5-p-tolyl-1,6-dihydropyrimidin-4-yl)benzonitrile,HCl salt

To a solution of tert-butyl1-(4-(4-cyanophenyl)-1-methyl-6-oxo-5-p-tolyl-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate (50 mg, 0.1 mmol) in EA (10 mL)was added a 4N HCl solution in EA (5 mL) and the mixture was stirred atRT for 2 h. The solvent was concentrated in vacuo, and the residue waspurified by preparative HPLC to give 20 mg (46%) of the title compoundas the hydrochloride salt. ¹H NMR (400 MHz, CDCl₃): δ 1.74-1.79 (m, 2H),2.00-2.04 (m, 2H), 2.21 (s, 3H), 2.96-3.03 (m, 2H), 3.29-3.03 (m, 1H),3.48 (s, 3H), 3.71-3.74 (m, 2H), 6.89 (d, J=8.0 Hz, 2H), 6.99 (d, J=8.0Hz, 2H), 7.38 (d, J=8.0 Hz, 2H), 7.44 (d, J=8.4 Hz, 2H). [M+H] Calc'dfor C₂₄H₂₅N₅O, 400; Found, 400.

Example 2:4-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 5% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.74-1.78 (m, 2H), 2.00-2.03 (m,2H), 2.98-3.02 (m, 2H), 3.26-3.00 (m, 1H), 3.48 (s, 3H), 3.69 (s, 3H),3.70-3.73 (m, 2H), 6.72 (d, J=8.8 Hz, 2H), 6.93 (d, J=8.4 Hz, 2H), 7.39(d, J=8.0 Hz, 2H), 7.46 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₄H₂₅N₅O₂,416; Found, 416.

Example 3:4-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 11% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.87-1.95 (m, 2H), 2.14-2.17 (m,2H), 3.15-3.24 (m, 2H), 3.43-3.48 (m, 1H), 3.62 (s, 3H), 3.93-3.98 (m,2H), 4.23 (s, 3H), 7.46 (d, J=9.2 Hz, 1H), 7.63 (d, J=8.0 Hz, 2H), 7.71(d, J=8.4 Hz, 2H), 8.12 (dd, J=8.8, 1.6 Hz, 1H), 8.28 (d, J=2.0 Hz, 1H).[M+H] Calc'd for C₂₃H₂₄N₆O₂, 417; Found, 417.

Example 4:4-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 4% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.79-1.80 (m, 2H), 2.03-2.05 (m,2H), 2.66 (s, 3H), 3.04-3.09 (m, 2H), 3.30-3.34 (m, 1H), 3.50 (s, 3H),3.83-3.88 (m, 2H), 7.48 (d, J=8.4 Hz, 2H), 7.58 (d, J=8.4 Hz, 2H), 7.64(d, J=8.4 Hz, 1H), 8.00 (dd, J=8.4, 2.0 Hz, 1H), 8.54 (d, J=8.0 Hz, 1H).[M+H] Calc'd for C₂₃H₂₄N₆O, 401; Found, 401.

Example 5:4-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 7% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.89-1.95 (m, 2H), 2.15-2.18 (m,2H), 3.14-3.18 (m, 2H), 3.44-3.46 (m, 1H), 3.60 (s, 3H), 3.88-3.90 (m,5H), 6.79 (d, J=8.4 Hz, 1H), 6.96-7.02 (m, 2H), 7.54 (d, J=8.0 Hz, 2H),7.64 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₄H₂₄FN₅O₂, 434; Found, 434.

Example 6:4-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

The title compound was prepared as the hydrochloride salt in 5% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.83-1.89 (m, 2H), 2.10-2.13 (m,2H), 3.05-3.11 (m, 2H), 3.35-3.38 (m, 1H), 3.55 (s, 3H), 3.76 (s, 3H),3.77-3.82 (m, 2H), 6.84 (d, J=8.8 Hz, 2H), 7.04 (d, J=8.8 Hz, 2H), 7.21(d, J=8.0 Hz, 1H), 7.35 (d, J=8.0 Hz, 1H), 7.53-7.56 (m, 1H). [M+H]Calc'd for C₂₄H₂₄FN₅O₂, 434; Found, 434.

Preparation 7A: tert-butyl1-(5-chloro-4-(3-fluoro-4-cyanophenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate

A mixture ofN-[1-(5,6-dichloro-3-methyl-4-oxo(3-hydropyrimidin-2-yl))(4-piperidyl)](tert-butoxy)carboxamide(150 g, 0.40 mol), 3-fluoro-4-cyanophenylboronic acid (65.8 g, 0.40mol), Pd(Ph₃P)₄ (9.3 g, 8 mmol) and 0.4 N Na₂CO₃ (2 L, 0.80 mol) in ACN(4 L) was flushed with nitrogen and stirred at 85° C. for 2 h. Water wasadded and the mixture was extracted with EA (3×). The organics werecombined, washed with water, washed with brine, dried and concentratedin vacuo. The residue was purified by silica chromatography (1:1, EA:PE)to give 95 g (57%) of the title compound. ¹H NMR (400 MHz, CDCl₃): δ1.45 (s, 9H), 1.54-1.61 (m, 2H), 2.05-2.13 (m, 2H), 2.99-3.08 (m, 2H),3.53-3.58 (s, 5H), 3.70 (s, 1H), 4.54 (d, J=6.0 Hz, 1H), 7.68-7.80 (m,3H).

Preparation 7B: tert-butylN-[1-[4-(4-cyano-3-fluorophenyl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxopyrimidin-2-yl]piperidin-4-yl]carbamate

A mixture of(tert-butoxy)-N-{1-[5-chloro-6-(4-cyano-3-fluorophenyl)-3-methyl-4-oxo(3-hydropyrimidin-2-yl)](4-piperidyl)}carboxamide(1 g, 2.169 mmol), 3-fluoro-4-methoxy benzeneboronic acid (740 mg, 4.338mmol), Pd(dppf)Cl₂ (480 mg, 0.651 mmol) and Na₂CO₃ (690 mg, 6.51 mmol)in dioxane:H₂O (3:1, 15 mL) was flushed with nitrogen, capped andstirred at 145° C. for 2 h in the microwave. The reaction mixture wasconcentrated and the residue was purified by FC (1:1, EA:PE) to give 800mg (71%) of the title compound. [M+H] Calc'd for C₂₉H₃₁F₂N₅O₄, 552;Found, 552. ¹H NMR (400 MHz, CDCl₃): δ ppm 1.46 (s, 9H), 1.60 (d,J=10.11 Hz, 2H), 2.11 (d, J=11.62 Hz, 2H), 3.06 (t, J=12.00 Hz, 2H),3.54 (s, 3H), 3.60 (d, J=13.64 Hz, 2H), 3.72 (br. s., 1H), 3.88 (s, 3H),4.52 (br. s., 1H), 6.79-6.89 (m, 2H), 6.97 (d, J=12.38 Hz, 1H), 7.13 (d,J=8.34 Hz, 1H), 7.31 (d, J=9.85 Hz, 1H), 7.42 (br. s., 1H).

Example 7:4-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a solution of tert-butylN-[1-[4-(4-cyano-3-fluorophenyl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxopyrimidin-2-yl]piperidin-4-yl]carbamate(5.2 g, 9.44 mmol) in EA (20 mL) was added a 1N HCl in EA (30 mL). Themixture was stirred at RT for 2 h. The solvent was concentrated in vacuoto give the title product as the HCl salt (4.05 g, 88%). ¹H NMR (400MHz, CD₃OD): δ 1.77-1.79 (m, 2H), 2.02-2.04 (m, 2H), 2.99-3.04 (m, 2H),3.26-3.00 (m, 1H), 3.38 (s, 3H), 3.73 (s, 3H), 3.73-3.75 (m, 2H),6.67-6.68 (m, 1H), 6.84-6.95 (m, 2H), 7.12-7.14 (m, 1H), 7.24-7.36 (m,1H), 7.46-7.50 (m, 1H). [M+H] Calc'd for C₂₄H₂₃F₂N₅O₂, 452; Found, 452.

Example 8:4-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

The title compound was prepared as the hydrochloride salt in 6% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.79-1.83 (m, 2H), 2.02-2.06 (m,2H), 3.04-3.11 (m, 2H), 3.21-3.22 (m, 1H), 3.49 (s, 3H), 3.81-3.85 (m,2H), 4.12 (s, 3H), 7.22-7.24 (m, 1H), 7.38 (d, J=9.2 Hz, 1H), 7.49 (d,J=9.2 Hz, 1H), 7.57-7.61 (m, 1H), 8.04-8.07 (m, 1H), 8.21 (s, 1H). [M+H]Calc'd for C₂₃H₂₃FN₆O₂, 435; Found, 435.

Example 9:4-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

The title compound was prepared as the hydrochloride salt in 8% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.92-1.96 (m, 2H), 2.16-2.19 (m,2H), 2.80 (s, 3H), 3.19-3.25 (m, 2H), 3.45-3.49 (m, 1H), 3.62 (s, 3H),3.96-3.99 (m, 2H), 7.34 (d, J=8.0 Hz, 1H), 7.60 (d, J=7.2 Hz, 1H), 7.71(t, J=7.6 Hz, 1H), 7.80 (d, J=8.4 Hz, 1H), 8.18 (d, J=8.4 Hz, 1H), 8.71(s, 1H). [M+H] Calc'd for C₂₃H₂₃FN₆O, 419; Found, 419.

Example 10:4-[2-(4-amino-piperidin-1-yl)-5-(6-ethyl-pyridin-3-yl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 7% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.30 (t, J=4.0 Hz, 3H), 1.83-1.88(m, 2H), 2.06-2.09 (m, 2H), 2.96-2.99 (m, 2H), 3.09-3.16 (m, 2H),3.26-3.31 (m, 1H), 3.51 (s, 3H), 3.86-3.89 (m, 2H), 7.35 (d, J=8.0 Hz,2H), 7.61 (d, J=8.0 Hz, 2H), 7.71 (d, J=8.4 Hz, 1H), 8.08 (d, J=8.4 Hz,1H), 8.57 (s, 1H). [M+H] Calc'd for C₂₄H₂₆N₆O, 415; Found, 415.

Example 11:2-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(4-methylamino-piperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 7% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.80-1.90 (m, 2H), 2.19-2.23 (m,2H), 2.75 (s, 3H), 3.06-3.12 (m, 2H), 3.32-3.36 (m, 1H), 3.56 (s, 3H),3.76 (s, 3H), 3.84-3.87 (m, 2H), 6.84 (d, J=8.4 Hz, 2H), 7.04 (d, J=8.4Hz, 2H), 7.22 (d, J=8.0 Hz, 1H), 7.36 (d, J=10.8 Hz, 1H), 8.54-7.58 (m,1H). [M+H] Calc'd for C₂₅H₂₆FN₅O₂, 448; Found, 448.

Example 12:2-fluoro-4-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-2-(4-methylamino-piperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile

The title compound was prepared as the hydrochloride salt in 7% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CD₃OD): δ 1.78-1.88 (m, 2H), 2.17-2.20 (m,2H), 2.73 (s, 3H), 3.05-3.11 (m, 2H), 3.30-3.35 (m, 1H), 3.54 (s, 3H),3.82 (s, 3H), 3.83-3.86 (m, 2H), 6.76 (d, J=8.4 Hz, 1H), 6.93-6.99 (m,2H), 7.20 (d, J=8.4 Hz, 1H), 7.38 (d, J=10.4 Hz, 1H), 8.55-7.589 (m,1H). [M+H] Calc'd for C₂₅H₂₅F₂N₅O₂, 466; Found, 466.

Preparation 13A: 2,6-dichloro-3-ethyl-3H-pyrimidin-4-one

A solution of 2,6-dichloro-pyrimidin-4-ol (1.0 g, 6.1 mmol) and K₂CO₃(1.1 g, 7.9 mmol) in DMF (10 mL) was stirred at RT for 15 min. Thereaction mixture was cooled to 0° C., and iodoethane (1.1 mL, 6.7 mmol)was added dropwise. After stirring overnight at RT, the reaction mixturewas diluted with EA, washed with brine, dried (Na₂SO₄) and concentratedin vacuo. The residue was purified by silica chromatography (20:1,EA:PE) to give 330 mg (28%) of the title compound. ¹H NMR (400 MHz,CDCl₃): δ 1.37 (t, J=7.6 Hz, 3H), 4.76 (q, J=6.8 Hz, 2H), 6.67 (s, 1H).[M+H] Calc'd for C₆H₆Cl₂N₂O, 193, 195, 197; Found, 193, 195, 197.

Preparation 13B:[1-(4-chloro-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-2-yl)-piperidin-4-yl]-carbamicacid tert-butyl ester

A solution of 2,6-dichloro-3-ethyl-3H-pyrimidin-4-one (320 mg, 1.64mmol), DIEA (423 mg, 3.28 mmol) and(tert-butoxy)-N-(4-piperidyl)carboxamide (328 mg, 1.64 mmol) in DMF (10mL) was heated to 120° C. for 1 h. The solvent was concentrated in vacuoand the residue was purified by silica chromatography (1:5, EA:PE) togive 210 mg (36%) of the title compound as a yellow solid. ¹H NMR (400MHz, CDCl₃): δ 1.25-1.32 (m 2H), 1.35 (t, J=7.2 Hz, 3H), 1.96-2.02 (m,2H), 2.98-3.06 (m, 2H), 3.70 (br, 1H), 4.30 (q, J=5.2 Hz, 2H), 4.44 (br,1H), 4.57-4.61 (m, 2H), 5.95 (s, 1H). [M+H] Calc'd for C₁₆H₂₅C₁N₄O₃,357, 359; Found, 357, 359.

Preparation 13C:{1-[4-(4-cyano-3-fluoro-phenyl)-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of[1-(4-chloro-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-2-yl)-piperidin-4-yl]-carbamicacid tert-butyl ester (210 mg, 0.59 mmol) in CH₃CN (10 mL),3-fluoro-4-cyanophenylboronic acid (126 mg, 0.77 mmol), Pd(PPh)₄ (14 mg,0.012 mmol) and 0.4 M Na₂CO₃ (4.5 mL, 1.77 mmol) was stirred at 90° C.overnight under N₂ atmosphere. The organic was concentrated in vacuo,and the aqueous extracted with DCM (2×). The combined organics werewashed with brine, dried (Na₂SO₄) and concentrated. The residue waspurified by silica chromatography (1:2, EA:PE) to give 185 mg (64%) ofthe title compound as a yellow solid. [M+H] Calc'd for C₂₃H₂₈FN₅O₃, 442;Found, 442.

Example 13:4-[2-(4-amino-piperidin-1-yl)-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a mixture of{1-[4-(4-cyano-3-fluoro-phenyl)-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (180 mg, 0.41 mmol) in EA (5 mL) was added a 4 Msolution of HCl in EA (3 mL). The reaction mixture was stirred for 30min. The solvent was evaporated in vacuo to give 150 mg of the titledcompound (97%) as a yellow solid (HCl salt). ¹H NMR (400 MHz, CD₃OD): δ1.28 (t, J=7.2 Hz, 1H), 1.48-1.52 (m, 2H), 1.99-2.02 (m, 2H), 2.94-3.01(m, 2H), 3.33-3.38 (m, 1H), 6.81 (q, J=6.8 Hz, 2H), 4.85-4.88 (m, 2H),6.95 (s, 1H), 7.73 (t, J=8.0 Hz, 1H), 7.90-7.95 (m, 2H). [M+H] Calc'dfor C₁₈H₂₀FN₅O, 342; Found, 342.

Preparation 14A:{1-[4-(4-cyano-3-fluoro-phenyl)-5-cyclopentylethynyl-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of tert-butyl1-(5-chloro-4-(3-fluoro-4-cyanophenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)piperidin-4-ylcarbamate(200 mg, 0.43 mmol), ethynyl-cyclopentane (82 mg, 0.87 mmol),Pd(MeCN)₂Cl₂ (4.5 mg, 0.017 mmol), X-Phos (10 mg, 0.022 mmol) and K₂CO₃(120 mg, 0.87 mmol) in ACN (15 mL) was stirred overnight at 95° C. in asealed tube. The reaction mixture was cooled to RT and the solvent wasconcentrated in vacuo. The residue was purified by silica chromatography(1:2, EA:PE) to give 100 mg (45%) of the title compound. [M+H] Calc'dfor C₂₉H₃₄FN₅O₃, 519; Found, 519.

Example 14:4-[2-(4-amino-piperidin-1-yl)-5-cyclopentylethynyl-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

The title compound was prepared as the hydrochloride salt in 70% overallyield according to the general procedure for the preparation ofExample 1. ¹H NMR (400 MHz, CDCl₃): δ 1.50-1.74 (m, 8H), 1.94-1.99 (m,4H), 2.88-3.01 (m, 4H), 3.51 (s, 3H), 3.60 (d, J=13.2 Hz, 2H), 7.63-7.67(m, 1H), 8.07-8.11 (m, 2H). [M+H] Calc'd for C₂₄H₂₆FN₅O, 419; Found,419.

Preparation 15A: (2,4,5-trichloro-6-oxo-6H-pyrimidin-1-yl)-acetic acidmethyl ester

To a solution of 2,5,6-trichloro-3H-pyrimidin-4-one (20.0 g, 0.1 mol) inDMF (150 mL) was added NaH (60% in mineral oil, 6.0 g, 0.12 mol) inportions at 0° C. and the mixture was stirred for 30 min. Bromoaceticacid methyl ester (18.3 g, 0.12 mol) was then added, and the reactionmixture was stirred at RT overnight. The solution was diluted with water(800 mL) and extracted with EA (200 mL, 3×). The combined organics werewashed with water (800 mL, 3×), washed with brine (500 mL), dried(Na₂SO₄) and concentrated. The residue was purified by silicachromatography (1:50, EA:PE) to give 6.0 g of the title product (22%).¹H NMR (400 MHz, CDCl₃): δ 3.80 (s, 3H), 5.04 (s, 2H). [M+H] Calc'd forC₇H₅C₁₃N₂O₃, 271; Found, 271.

Preparation 15B:[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4,5-dichloro-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester

To a solution of (2,4,5-trichloro-6-oxo-6H-pyrimidin-1-yl)-acetic acidmethyl ester (6.0 g, 22.4 mmol) and piperidin-4-yl-carbamic acidtert-butyl ester (4.9 g, 24.4 mmol) in DMF (50 mL) was added DIPEA (5.7g, 44.3 mmol) dropwise at RT, and the mixture was stirred overnight. Thereaction mixture was diluted with water (500 mL), and the solids werecollected by filtration. The solids were then dissolved in DCM (100 mL),washed with water (100 mL, 3×), washed with brine (100 mL), dried(Na₂SO₄) and concentrated. The residue was purified by silicachromatography (1:2 to 1:1, DCM:PE) to give 6.3 g of the title product(64%). ¹H NMR (400 MHz, CDCl₃): δ 1.22-1.34 (m, 2H), 1.45 (s, 9H),1.97-2.03 (m, 2H), 2.96-3.09 (m, 2H), 3.68-3.69 (m, 1H), 3.75 (s, 3H),4.42-4.44 (m, 3H), 4.84 (s, 2H). [M+H] Calc'd for C₁₇H₂₄Cl₂N₄O₅, 435;Found, 435.

Preparation 15C:[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-5-chloro-4-(4-cyano-3-fluoro-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester

A mixture of[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4,5-dichloro-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester (5.76 g, 13.2 mmol), 4-cyano-3-fluoro benzeneboronicacid (2.24 g, 16.1 mmol), Pd(PPh₃)₄ (306 mmol, 0.26 mmol) and Na₂CO₃(2.8 g, 26.5 mmol) in DMF:H₂O (50 mL: 10 mL) was stirred at 65° C.overnight under nitrogen atmosphere. The reaction mixture wasconcentrated, and the residue was purified by silica chromatography(1:20 to 1:0, EA:PE) to give 2.4 g of the title product (43%). ¹H NMR(400 MHz, CDCl₃): δ 1.27-1.37 (m, 2H), 1.45 (s, 9H), 1.99-2.02 (m, 2H),2.99-3.06 (m, 2H), 3.68-3.76 (m, 1H), 3.78 (s, 3H), 4.42-4.52 (m, 3H),4.90 (s, 2H), 7.63-7.66 (m, 1H), 7.67-7.71 (m, 2H). [M+H] Calc'd forC₂₄H₂₇C₁FN₅O₅, 520; Found, 520.

Preparation 15D:[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester

A solution of[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-5-chloro-4-(4-cyano-3-fluoro-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester (2.2 g, 4.2 mmol), p-methoxyboronic acid (1.9 g, 12.7mmol), Pd-118 (274 mg, 0.42 mmol) and K₂CO₃ (1.2 g, 8.4 mmol) in DMF (50mL) was stirred at 145° C. for 6 h under nitrogen atmosphere. Thereaction mixture was diluted with water and extracted with EA (3×). Thecombined organics were washed with water, washed brine, dried (Na₂SO₄)and concentrated. The residue was purified by preparative HPLC to give600 mg of the title product (24%). [M+H] Calc'd for C₃₁H₃₄FN₅O₆, 592;Found, 592.

Preparation 15E:4-[2-(4-amino-piperidin-1-yl)-1-cyclopropylmethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a solution of[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid methyl ester (600 mg, 1.02 mmol) in MeOH (10 mL) was added a 2NNaOH solution (5 mL). After completion of the reaction, the solution wasacidified with 1N HCl and extracted with EA (3×). The combined organicswere washed with brine, dried (Na₂SO₄) and concentrated. The residue waspurified by preparative HPLC to give 240 mg of the title product as ayellow solid (41%). [M+H] Calc'd for C₃₀H₃₂FN₅O₆, 578; Found, 578.

Example 15:[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid

To a solution of[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid (100 mg, 0.15 mmol) in EA (10 mL) was added a 5N HCl solution in EA(5 mL). The reaction mixture was stirred at RT for 2 h, and the solventwas concentrated in vacuo. The residue was purified by preparative HPLCto give 25 mg of the title product as HCl salt (32%). ¹H NMR (400 MHz,CD3OD): δ 1.53-1.56 (m, 2H), 2.00-2.03 (m, 2H), 3.00-3.07 (m, 2H),3.35-3.39 (m, 1H), 3.67 (s, 3H), 4.70 (s, 2H), 4.76-4.77 (m, 2H), 6.74(d, J=8.4 Hz, 2H), 6.96 (d, J=8.8 Hz, 2H), 7.17 (d, J=8.4 Hz, 1H), 7.26(d, J=10.0 Hz, 1H), 7.50 (dd, J=7.2, 8.0 Hz, 1H). [M+H] Calc'd forC₂₅H₂₄FN₅O₄, 478; Found, 478.

Preparation 16A:{1-[1-carbamoylmethyl-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

To a solution of[2-(4-tert-butoxycarbonylamino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-aceticacid (120 mg, 0.2 mmol) in DMF (5 mL) was added NH₄Cl (17 mg, 0.3 mmol),HATU (95 mg, 0.25 mmol) and DIEA (25 mg, 0.4 mmol). After completion ofthe reaction, the solution was diluted with H₂O and extracted with DCMfor (3×). The combined organics were dried (Na₂SO₄) and concentrated.The residue was purified by preparative HPLC to give 50 mg of the titleproduct as a yellow solid (43%). [M+H] Calc'd for C₃₀H₃₃FN₆O₅, 577;Found, 577.

Example 16:2-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-acetamide

The title compound was prepared as the hydrochloride salt in 96% yieldaccording to the procedure for the preparation of Example 15. ¹H NMR(400 MHz, CD₃OD): δ 1.49-1.53 (m, 2H), 1.98-2.01 (m, 2H), 2.97-3.04 (m,2H), 3.33-3.36 (m, 1H), 3.68 (s, 3H), 4.69 (s, 2H), 4.75-4.78 (m, 2H),6.75 (d, J=8.4 Hz, 2H), 6.99 (d, J=8.8 Hz, 2H), 7.16 (dd, J=1.2, 8.0 Hz,1H), 7.25 (dd, J=0.8, 10.4 Hz, 1H), 7.49 (dd, J=7.2, 8.0 Hz, 1H). [M+H]Calc'd for C₂₅H₂₅FN₆O₃, 477; Found, 477.

Preparation 17A: 2,6-dichloro-3-(3-methoxy-propyl)-3H-pyrimidin-4-one

To a solution of 2,6-dichloro-3H-pyrimidin-4-one (600 mg, 3.65 mmol) inDMF (10 mL) was added K₂CO₃ (1.0 g, 7.3 mmol) and the mixture wasstirred at RT for 10 min. 1-Bromo-3-methoxy-propane (101 mg, 7.3 mmol)was then added dropwise at 0° C., and the mixture was stirred at RTovernight. DMF was concentrated in vacuo, and the residue was purifiedby silica chromatography to give 400 mg of the title compound (47%).[M+H] Calc'd for; Calc'd for C₈H₁₀Cl₂N₂O₂, 237; Found, 237.

Preparation 17B:{1-[4-chloro-1-(3-methoxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A solution of 2,6-dichloro-3-(3-methoxy-propyl)-3H-pyrimidin-4-one (400mg, 1.68 mmol), piperidin-4-yl-carbamic acid tert-butyl ester (405 mg, 2mmol) and DIEA (260 mg, 2.0 mmol) in DMF (20 mL) was stirred at 85° C.for 2 h. The solvent was concentrated, and the residue was purified bysilica chromatography to give 500 mg of the title compound (75%). [M+H]Calc'd for C₁₈H₂₉C₁N₄O₄, 400; Found, 400.

Preparation 17C:{1-[4-(4-cyano-3-fluoro-phenyl)-1-(3-methoxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of{1-[4-chloro-1-(3-hydroxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (200 mg, 0.5 mmol), 4-cyano-3-fluorophenyl boricacid (107 mg, 0.65 mmol), Pd(PPh₃)₄ (12 mg, 0.01 mmol) and 0.4M Na₂CO₃solution (4 mL) in ACN was stirred at 85° C. overnight. The reactionmixture was diluted with water and extracted with EA (3×). The reactionmixture was stirred at RT for 2 h and the solvent was concentrated invacuo. The residue was purified by silica chromatography to give 240 mgof the title product (99%). [M+H] Calc'd for C₂₅H₃₂FN₅O₄, 485; Found,485.

Example 17:4-[2-(4-amino-piperidin-1-yl)-1-(3-hydroxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a solution of{1-[4-(4-cyano-3-fluoro-phenyl)-1-(3-methoxy-propyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (200 mg, 0.41 mmol) in DCM was added 1M BBr₃ (4mL) at −78° C. The mixture was stirred at RT for 2 h and quenched at 0°C. with MeOH. The solution was washed with aqueous saturated NaHCO₃. Theorganic layer was dried and concentrated. The residue was purified bypreparative HPLC to give 35 mg of the title product as the hydrochloridesalt (23%). ¹H NMR (400 MHz, CD₃OD): 1.65-1.69 (m, 2H), 1.97-2.19 (m,4H), 3.13-3.22 (m, 2H), 3.48-3.55 (m, 1H), 3.73 (t, J=8.0 Hz, 2H), 4.55(t, J=8.0 Hz, 2H), 4.94-4.95 (m, 2H), 6.71 (s, 1H), 7.88-8.05 (m, 3H).[M+H] Calc'd for C₁₉H₂₂FN₅O₂, 371; Found, 371.

Preparation 18A:{1-[5-benzofuran-5-yl-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of{1-[5-chloro-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (200 mg, 0.45 mmol), benzofuran-5-boronic acid(120 mg, 0.68 mmol), Pd(PPh₃)₄ (26 mg, 0.05 mmol) and 2M Na₂CO₃ (0.9 mL)in 1,4-dioxane (200 mL) was refluxed overnight under N₂ atmosphere. Thereaction mixture was diluted with water and extracted with EA (3×). Thecombined organics were washed with brine, dried (Na₂SO₄) andconcentrated. The residue was purified by silica chromatography to give100 mg of the title product (42%). [M+H] Calc'd for C₃₀H₃₀FN₅O₄, 543;Found, 543.

Example 18:4-[2-(4-amino-piperidin-1-yl)-5-benzofuran-5-yl-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a solution of Preparation 18A (60 mg, 0.11 mmol) in EA (20 mL) wasadded a 4M HCl solution in EA (10 mL). The mixture was stirred at RT for2 h. The solvent was concentrated in vacuo to give 43 mg of the titleproduct as the hydrochloride salt (53%). ¹H NMR (400 MHz, CD₃OD):1.85-1.92 (m, 2H), 2.13-2.18 (m, 2H), 3.10 (t, J=4.0 Hz, 2H), 3.31-3.33(m, 1H), 3.61 (s, 3H), 3.87 (d, J=13.2 Hz, 2H), 6.65-7.21 (m, 3H),7.38-7.76 (m, 4H), 7.76 (s, 1H). [M+H] Calc'd for C₂₅H₂₂FN₅O₂, 443;Found, 443.

Preparation 19A:{1-[5-cyano-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of{1-[5-chloro-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (460 mg, 1 mmol), Zn(CN)₂ (175 mg, 1.5 mmol) andPd(PPh₃)₄ (116 mg, 0.0.1 mmol) in DMF (5 mL) was stirred for 4 h at 150°C. under N₂ atmosphere. The reaction mixture was cooled to RT andfiltered. The filtrate was concentrated in vacuo, and the residue waspurified by preparative HPLC to give 150 mg of the title product as ayellow solid (33%). [M+H] Calc'd for C₂₃H₂₅FN₆O₃, 453; Found, 453.

Example 19:2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidine-5-carbonitrile

To a solution of{1-[5-cyano-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5NHCl solution in EA (5 mL). The reaction mixture was stirred at RT for 2h, and the solvent was concentrated in vacuo to give 120 mg of the titleproduct as HCl salt (94%). ¹H NMR (400 MHz, CD₃OD): δ 1.67-1.72 (m, 2H),2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H),3.98-4.02 (m, 2H), 7.82-7.90 (m, 3H). [M+H] Calc'd for C₁₈H₁₇FN₆O, 353;Found, 353.

Example 20:4-[2-(4-aminopiperidin-1-yl)-5-chloro-1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile

To a solution of{1-[5-chloro-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5NHCl solution in EA (5 mL). The reaction mixture was stirred at RT for 2h, and the solvent was concentrated in vacuo to give 120 mg of the titleproduct as HCl salt (94%). ¹H NMR (400 MHz, CD₃OD): δ 1.67-1.72 (m, 2H),2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H),3.98-4.02 (m, 2H), 7.82-7.90 (m, 3H). [M+H] Calc'd for C₁₈H₁₇FN₆O, 353;Found, 353. ¹H NMR (400 MHz, METHANOL-d₄): δ ppm 1.73-1.91 (m, 2H), 2.18(d, J=12.13 Hz, 2H), 3.06 (t, J=12.76 Hz, 2H), 3.33-3.40 (m, 1H), 3.57(s, 3H), 3.83 (d, J=13.14 Hz, 2H), 7.75-7.93 (m, 3H).

TABLE 3 Chemical MS Synthesis Structure (ESI) Example (prepared byprocedure of cited Example) m/z NMR spectrum data 21

433 ¹H NMR (400 MHz, CD₃OD): δ 1.89-1.93 (m, 2H), 2.18-2.21 (m, 2H),2.73 (s, 3H), 2.74 (s, 3H), 3.11-3.17 (m, 2H), 3.33-3.39 (m, 1H), 3.57(s, 3H), 3.4-3.97 (m, 2H), 7.28 (d, J = 8.0 Hz, 1H), 7.55 (d, J = 10.0Hz, 1H), 7.63- 7.67 (m, 1H), 7.74 (d, J = 8.4 Hz, 1H), 8.12 (d, J = 8.4Hz, 1H), 8.65 (s, 1H). 22

492 ¹H NMR (400 MHz, CDCl₃): δ 1.74-1.80 (m, 4H), 1.93-1.97 (m, 2H),3.11 (s, 2H), 3.26-3.35 (m, 6H), 3.47 (s, 3H), 3.75 (s, 3H), 6.68 (dd, J= 1.2, 8.4 Hz, 1H), 6.86-6.72 (m, 2H), 7.11 (dd, J = 1.2, 8.0 Hz, 1H),7.30 (dd J = 1.2, 10.8 Hz, 1H), 7.46 (dd J = 6.8, 7.6 Hz, 1H). 23

472 ¹H NMR (400 MHz, CD₃OD): 1.75-1.82 (m, 2H), 2.03-2.06 (m, 2H),3.06-3.12 (m, 2H), 3.22-3.34 (m, 1H), 3.49 (s, 3H), 3.1 (d, J = 13.6 Hz,2H), 7.07-7.09 (m, 1H), 7.36-7.38 (m, 1H), 7.51-7.55 (m, 1H), 7.66 (d, J= 8.0 Hz, 1H), 7.79-7.82 (m, 1H), 8.33 (s, 1H). 24

439 ¹H NMR (400 MHz, CD₃OD): 1.96-2.01 (m, 2H), 2.20-2.22 (m, 2H),3.23-3.32 (m, 2H), 3.46-3.49 (m, 1H), 3.65 (s, 3H), 3.94-3.97 (m, 2H),4.39 (s, 3H), 7.55-7.77 (m, 7H), 8.76 (s, 1H). 25

452 ¹H NMR (300 MHz, CD₃OD): δ 1.72-1.93 (m, 3H), 1.97-2.23 (m, 1H),3.16-3.30 (m, 2H), 3.50- 3.55 (m, 2H), 3.60 (s, 3H), 3.83- 3.84 (m, 1H),3.86 (s, 3H), 6.82 (d, J = 8.1 Hz, 1H), 6.97-7.05 (m, 2H), 7.25 (d, J =8.1 Hz, 1H), 7.44 (d, J = 10.8 Hz, 1H), 7.62 (t, J = 7.5 Hz, 1H). 26

453 ¹H NMR (400 MHz, CD₃OD): 1.64-1.69 (m, 2H), 1.89-1.92 (m, 2H),2.85-2.91 (m, 2H), 3.15-3.20 (m, 1H), 3.34 (s, 3H), 3.62 (d, J = 8.4 Hz,2H), 3.71 (s, 3H), 6.99 (d, J = 8.4 Hz, 1H), 7.20-7.40 (m, 4H). 27

438 ¹H NMR (400 MHz, CD₃OD): δ 2.19-2.22 (m, 1H), 2.49-2.51 (m, 1H),3.63 (s, 3H), 3.75-3.81 (m, 2H), 3.87 (s, 3H), 3.87-3.93 (m, 1H),4.02-4.06 (m, 2H), 6.80 (d, J = 8.4 Hz, 1H), 7.00 (t, J = 10.8 Hz, 2H),7.25 (d, J = 9.6 Hz, 1H), 7.44 (d, J = 10.8 Hz, 1H), 7.61 (t, J = 7.4Hz, 28

452 ¹H NMR (400 MHz, CD₃OD): δ 1.69-1.99 (m, 3H), 2.14-2.19 (m, 1H),3.09-3.24 (m, 2H), 3.43- 3.46 (m, 1H), 3.56-3.60 (m, 4H), 3.77-3.80 (m,1H), 3.82 (s, 3H), 6.77 (d, J = 8.0 Hz, 1H), 6.94- 7.00 (m, 2H), 7.21(d, J = 8.0 Hz, 1H), 7.39 (d, J = 10.4 Hz, 1H), 8.56-7.60 (m, 1H). 29

438 ¹H NMR (400 MHz, CD₃OD): δ 2.25-2.29 (m, 1H), 2.50-2.55 (m, 1H),3.69 (s, 3H), 3.89-3.84 (m, 5H), 3.99-4.03 (m, 1H), 5.05- 4.16 (m, 2H),6.80 (d, J = 8.4 Hz, 1H), 6.97-7.03 (m, 2H), 7.29 (dd, J = 2.4, 8.0 Hz,1H), 7.47 (d, J = 10.4 Hz, 1H), 7.64 (dd, J = 6.8, 8.0 Hz, 1H). 30

434 ¹H NMR (400 MHz, CD₃OD): δ 1.73-2.02 (m, 3H), 2.19-2.23 (m, 1H),3.13-3.26 (m, 2H), 3.49- 3.52 (m, 2H), 3.60 (s, 3H), 3.77- 3.85 (m, 1H),3.85 (s, 3H), 6.89 (d, J = 11.6 Hz, 2H), 7.08-7.10 (d, J = 11.6 Hz, 2H),7.24 (d, J = 8.0 Hz, 1H), 7.41 (d, J = 10.8 Hz, 1H), 7.57-7.61 (m, 1H).31

434 ¹H NMR (400 MHz, CD₃OD): δ 1.69-1.99 (m, 3H), 2.07-2.10 (m, 1H),3.09-3.24 (m, 2H), 3.43- 3.46 (m, 1H), 3.56-3.60 (m, 4H), 3.68 (s, 3H),3.76-3.79 (m, 1H), 6.75 (d, J = 8.8 Hz, 2H), 6.96 (d, J = 9.2 Hz, 2H),7.13 (dd, J = 2.0, 8.0 Hz, 1H), 7.27 (dd, J = 0.8, 10.4 Hz, 1H), 7.47(dd, J = 6.8, 8.0 Hz, 1H). 32

466 ¹H NMR (400 MHz, CD₃OD): δ 1.41 (s, 3H), 1.82-1.85 (m, 2H),1.91-1.99 (m, 2H), 3.22-3.25 (m, 2H), 3.47 (s, 3H), 3.50-3.57 (m, 2H),3.75 (s, 3H), 6.69 (d, J = 8.4 Hz, 1H), 6.86-6.92 (m, 2H), 7.14 (d, J =8.4 Hz, 1H), 7.32 (d, J = 10.8 Hz, 1H), 7.47-7.51 (m, 1H). 33

439 ¹H NMR (400 MHz, CD3OD): δ 1.95-1.99 (m, 2H), 2.19-2.22 (m, 2H),3.20-3.26 (m, 2H), 3.45- 3.50 (m, 1H), 3.63 (s, 3H), 3.90 (d, J = 12.8Hz, 2H), 4.06 (s, 3H), 7.21 (d, J = 8.4 Hz, 1H), 7.48- 7.57 (m, 6H),7.96 (s, 1H). 34

476 ¹H NMR (400 MHz, CD₃OD): δ 1.75-1.79 (m, 2H), 2.02-2.05 (m, 2H),3.00-3.06 (m, 2H), 3.21- 3.31 (m, 1H), 3.48 (s, 3H), 3.72- 3.75 (m, 2H),4.77-4.81 (m, 2H), 7.22 (s, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.39 (d, J =2.0 Hz, 1H), 7.60-7.64 (m, 2H). 35

457 ¹H NMR (400 MHz, CD₃OD): 1.85-1.99 (m, 2H), 2.18-2.20 (m, 2H),3.19-3.24 (m, 2H), 3.46-3.50 (m, 1H), 3.86 (s, 3H), 3.86-3.92 (m, 2H),4.10 (s, 3H), 7.21-7.25 (m, 2H), 7.40-7.53 (m, 4H), 8.01 (s, 1H). 36

458 ¹H NMR (400 MHz, CD₃OD): δ 1.79-1.82 (m, 2H), 2.04-2.07 (m, 2H),3.09-3.15 (m, 2H), 3.32- 3.38 (m, 1H), 3.50 (s, 3H), 3.76- 3.79 (m, 2H),4.74-4.78 (m, 2H), 7.12 (s, 1H), 7.55 (d, J = 8.4 Hz, 2H), 7.62 (s, 1H),7.67 (d, J = 8.0 Hz, 2H). 37

458 ¹H NMR (400 MHz, CD₃OD): δ 1.98-2.04 (m, 2H), 2.21-2.24 (m, 2H),3.27-3.30 (m, 2H), 3.50- 3.52 (m, 1H), 3.65 (s, 3H), 3.98 (d, J = 12.8Hz, 2H), 4.42 (s, 3H), 7.33 (d, J = 8.0 Hz, 1H), 7.49 (d, J = 10.0 Hz,1H), 7.60-7.73 (m, 3H), 7.84 (s, 1H), 8.85 (s, 1H). 38

452 ¹H NMR (400 MHz, CD₃OD): 1.87-1.94 (m, 2H), 2.15 (d, J = 12.0 Hz,2H), 3.13 (t, J = 8.4 Hz, 2H), 3.39-3.43 (m, 1H), 3.59 (s, 3H), 3.87 (d,J = 12.8 Hz, 2H), 3.97 (s, 3H), 6.79 (d, J = 8.8 Hz, 2H), 7.55 (d, J =8.4 Hz, 2H), 7.70 (d, J = 8.4 Hz, 2H). 39

448 ¹H NMR (400 MHz, CD₃OD): δ 1.91-1.94 (m, 2H), 2.16-2.19 (m, 2H),3.15-3.21 (m, 2H), 3.50- 3.52 (m, 1H), 3.61 (s, 3H), 3.90 (d, J = 12.4Hz, 2H), 7.22 (d, J = 8.0 Hz, 1H), 7.30 (d, J = 8.0 Hz, 2H), 7.43 (d, J= 10.8 Hz, 1H), 7.59 (t, J = 1.2 Hz, 1H), 7.86 (d, J = 8.0 Hz, 2H). 40

461 ¹H NMR (400 MHz, CD₃OD): δ 1.88-1.91 (m, 2H), 2.12-2.13 (m, 2H),2.94 (s, 3H), 3.13-3.15 (m, 2H), 3.30-3.34 (m, 1H), 3.61 (s, 3H), 3.89(d, J = 14.4 Hz, 2H), 7.00 (d, J = 8.0 Hz, 1H), 7.11 (d, J = 12.0 Hz,1H), 7.53 (d, J = 12.0 Hz, 2H), 7.61-7.64 (m, 3H). 41

447 ¹H NMR (400 MHz, CD₃OD): δ 1.87-1.91 (m, 2H), 2.14-2.16 (m, 2H),3.15 (t, J = 12.0 Hz, 2H), 3.30-3.40 (m, 1H), 3.61 (s, 3H), 3.89 (d, J =14.0 Hz, 2H), 7.01 (d, J = 8.0 Hz, 1H), 7.13 (d, J = 12.0 Hz, 1H),7.52-7.77 (m, 5H). 42

459 ¹H NMR (400 MHz, CD₃OD): δ 178-1.79 (m, 2H), 2.03-2.05 (m, 2H),3.00-3.06 (m, 2H), 3.21-3.31 (m, 1H), 3.49 (s, 3H), 3.75-3.78 (m, 2H),4.32 (s, 2H), 7.06 (dd, J = 1.2, 8.0 Hz, 1H), 7.12 (dd, J = 1.2, 8.0 Hz,1H), 7.31-7.36 (m, 2H), 7.42 (dd, J = 6.4, 7.6 Hz, 1H), 7.58 (d, J = 7.6Hz, 1H). 43

448 ¹H NMR (400 MHz, CD₃OD): δ 1.77-1.80 (m, 2H), 2.02-2.05 (m, 2H),3.01-3.05 (m, 2H), 3.35- 3.36 (m, 1H), 3.49 (s, 3H), 3.74- 3.98 (m, 2H),7.07 (dd, J = 1.6, 8.4 Hz, 1H), 7.27-7.32 (m, 3H), 7.44 (dd, J = 6.8,8.0 Hz, 1H), 7.73 (d, J = 1.2 Hz, 1H), 7.84 (d, J = 1.2 Hz, 1H). 44

434 ¹H NMR (400 MHz, CD₃OD): δ 1.87-1.91 (m, 1H), 2.25-2.28 (m, 1H),2.87-2.92 (m, 1H), 3.11- 3.17 (m, 1H), 3.30-3.32 (m, 1H), 3.41-3.56 (m,5H), 3.69-3.71 (m, 2H), 3.84 (s, 3H), 6.75 (d, J = 8.4 Hz, 1H),6.92-6.96 (m, 2H), 7.53 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.4 Hz, 2H).45

434 ¹H NMR (400 MHz, CD₃OD): δ 1.74-1.80 (m, 1H), 2.14-2.19 (m, 1H),2.77-2.81 (m, 1H), 3.01- 3.06 (m, 1H), 3.31-3.34 (m, 1H), 3.36-3.45 (m,5H), 3.59-3.60 (m, 2H) 3.71 (s, 3H), 6.63 (d, J = 8.4 Hz, 1H), 6.80-6.84(m, 2H), 7.44 (d, J = 8.0 Hz, 2H), 7.60 (d, J = 8.4 Hz, 2H). 46

452 ¹H NMR (400 MHz, CD₃OD): δ 2.15-2.18 (m, 2H), 3.31-3.34 (m, 2H),3.46-3.51 (m, 5H), 3.56- 3.59 (m, 2H), 3.74 (s, 3H), 3.78- 3.81 (m, 2H),6.68 (dd, J = 1.2, 8.4 Hz, 1H), 6.85-6.89 (m, 2H), 7.12 (dd, J = 1.2,7.6 Hz, 1H), 7.28 (dd, J = 1.6, 10.8 Hz, 1H), 7.47 (dd, J = 6.8, 8.0 Hz,1H). 47

438 ¹H NMR (400 MHz, DMSO-d₆): δ 3.27-3.34 (m, 4H), 3.45 (s, 3H),3.51-3.53 (m, 4H), 3.81 (s, 3H), 6.78 (d, J = 8.4 Hz, 1H), 7.02- 7.08(m, 2H), 7.18 (dd, J = 1.6, 8.4 Hz, 1H), 7.45 (dd, J = 1.6, 10.8 Hz,1H), 7.80 (dd, J = 7.2, 8.0 Hz, 1H), 9.41 (br, 1H). 48

434 ¹H NMR (400 MHz, CD₃OD): δ 184-1.94 (m, 2H), 2.20-2.23 (m, 2H),3.00-3.07 (m, 2H), 3.38-3.42 (m, 5H), 3.72 (s, 3H), 4.22-4.27 (m, 1H),6.61 (d, J = 8.8 Hz, 1H), 6.79-6.83 (m, 2H), 7.39 (d, J = 8.0 Hz, 2H),7.51 (d, J = 8.0 Hz, 2H). 49

449 ¹H NMR (400 MHz, CD₃OD): δ 1.82-1.87 (m, 2H), 2.04-2.07 (m, 2H),3.06-3.12 (m, 2H), 3.25 (s, 6H), 3.28-3.39 (m, 1H), 3.49 (s, 3H),3.81-3.84 (m, 2H), 7.37 (d, J = 8.0 Hz, 1H), 7.56 (d, J = 9.6 Hz, 1H),7.42 (t, J = 6.8 Hz, 1H), 8.31 (s, 2H). 50

462 ¹H NMR (400 MHz, CD₃OD): δ 1.93-1.97 (m, 2H), 2.17-2.20 (m, 2H),3.03 (s, 3H), 3.20-3.26 (m, 2H), 3.47-3.53 (m, 1H), 3.62 (s, 3H),3.98-4.02 (m, 2H), 7.32 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 10.0 Hz, 1H),7.67 (t, J = 6.4 Hz, 1H), 8.32 (s, 2H), 8.83 (s, 1H). 51

434 ¹H NMR (400 MHz, CD₃OD): δ 1.89-1.91 (m, 1H), 2.26-2.28 (m, 1H),2.91-2.93 (m, 1H), 3.12- 3.15 (m, 1H), 3.30-3.32 (m, 1H), 3.42-3.55 (m,5H), 3.70-3.72 (m, 2H) 3.84 (s, 3H), 6.84 (d, J = 8.4 Hz, 2H), 7.03 (d,J = 8.4 Hz, 2H), 7.29 (d, J = 8.0 Hz, 1H), 7.39 (d, J = 10.0 Hz, 1H),7.64 (t, J = 7.2 Hz, 1H). 52

434 ¹H NMR (400 MHz, CD₃OD): δ 1.86-1.91 (m, 1H), 2.22-2.28 (m, 1H),2.97-2.91 (m, 1H), 3.10- 3.13 (m, 1H), 3.29-3.32 (m, 1H), 3.40-3.51 (m,5H), 3.67-3.69 (m, 2H) 3.82 (s, 3H), 6.84 (d, J = 8.0 Hz, 2H), 7.02 (d,J = 8.4 Hz, 2H), 7.27 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 10.4 Hz, 1H),7.64 (t, J = 7.2 Hz, 1H). 53

434 ¹H NMR (400 MHz, CD₃OD): δ 2.03-2.06 (m, 2H), 2.32-2.35 (m, 2H),3.14-3.21 (m, 2H), 3.51- 3.56 (m, 5H), 3.78 (s, 3H), 4.37- 4.39 (m, 1H),6.84 (d, J = 7.2 Hz, 2H), 7.02 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 8.0 Hz,1H), 7.38 (d, J = 10.4 Hz, 1H), 7.62 (t, J = 7.2 Hz, 1H). 54

448 ¹H NMR (400 MHz, CD₃OD): δ 1.66-1.71 (m, 1H), 2.11-2.16 (m, 1H),2.77-2.81 (m, 1H), 2.93- 2.97 (m, 4H), 3.16-3.20 (m, 1H), 3.30-3.38 (m,2H), 3.43-3.50 (m, 5H), 3.69 (s, 3H), 6.75 (d, J = 8.4 Hz, 2H), 6.95 (d,J = 8.4 Hz, 2H), 7.16 (d, J = 8.4 Hz, 1H), 7.28 (d, J = 10.8 Hz, 1H),7.50 (dd, J = 6.8, 8.0 Hz, 1H). 55

448 ¹H NMR (400 MHz, CD₃OD): δ 2.03-2.13 (m, 4H), 2.84 (s, 3H),3.01-3.05 (m, 2H), 3.39-3.43 (m, 2H), 3.48 (s, 3H), 3.67 (s, 3H),3.87-3.92 (m, 1H), 6.74 (d, J = 8.8 Hz, 2H), 6.95 (d, J = 8.8 Hz, 2H),7.13 (dd, J = 1.2, 8.0 Hz, 1H), 7.21 (dd, J = 1.6, 10.4 Hz, 1H), 7.45(dd, J = 6.8, 7.6 Hz, 1H). 56

448 ¹H NMR (400 MHz, CD₃OD): δ 1.77-1.80 (m, 1H), 2.22-2.26 (m, 1H),2.90-2.92 (m, 1H), 3.03- 3.07 (m, 4H), 3.27-3.30 (m, 1H), 3.39-3.41 (m,2H), 3.44-3.46 (m, 5H), 3.77 (s, 3H), 6.86 (d, J = 8.4 Hz, 2H), 7.06 (d,J = 8.0 Hz, 2H), 7.29 (d, J = 8.4 Hz, 1H), 7.36 (d, J = 8.4 Hz, 1H),7.58 (dd, J = 6.8, 7.6 Hz, 1H). 57

448 ¹H NMR (400 MHz, CD₃OD): δ 1.77-1.85 (m, 2H), 2.03-2.06 (m, 2H),3.03-3.09 (m, 2H), 3.18 (s, 6H), 3.31-3.38 (m, 1H), 3.48 (s, 3H),3.78-3.81 (m, 2H), 7.03 (d, J = 9.2 Hz, 1H), 7.28 (d, J = 7.6 Hz, 1H),7.46 (d, J = 10.0 Hz, 1H), 7.59-7.63 (m, 2H), 7.71 (s, 1H). 58

449 ¹H NMR (400 MHz, CD₃OD): δ 1.83-1.88 (m, 2H), 2.21-2.24 (m, 2H),2.77 (s, 3H), 3.06-3.14 (m, 2H), 3.31-3.32 (m, 1H), 3.58 (s, 3H),3.87-3.91 (m, 5H), 6.83 (d, J = 11.2 Hz, 1H), 7.22 (dd, J = 2.0, 10.8Hz, 1H), 7.42 (dd, J = 2.0, 14.4 Hz, 1H), 7.59-7.65 (m, 2H), 7.84 (d, J= 3.2 Hz, 1H). 59

447 ¹H NMR (400 MHz, CD₃OD): δ 1.88-1.89 (m, 2H), 2.14-2.19 (m, 2H),3.13-3.19 (m, 2H), 3.28 (s, 6H), 3.41-3.46 (m, 1H), 3.60 (s, 3H),3.87-3.91 (m, 2H), 7.24 (d, J = 10.8 Hz, 1H), 7.39-7.44 (m, 3H),7.59-7.67 (m, 3H). 60

474 ¹H NMR (400 MHz, CD₃OD): δ 1.91-1.97 (m, 2H), 2.16-2.20 (m, 6H),3.14-3.20 (m, 2H), 3.47- 3.49 (m, 1H), 3.60-3.63 (m, 7H), 3.89-3.92 (m,2H), 7.31 (d, J = 9.6 Hz, 1H), 7.01 (d, J = 9.6 Hz, 1H), 7.41 (d, J =8.0 Hz, 1H), 7.58 (d, J = 10.0 Hz, 1H), 7.68- 7.75 (m, 2H), 7.79 (s,1H). 61

435 ¹H NMR (400 MHz, CD₃OD): δ 2.27-2.30 (m, 2H), 3.44-.347 (m, 2H),3.60-3.64 (m, 5H), 3.70- 3.73 (m, 2H), 3.91-3.94 (m, 5H), 6.83 (d, J =8.4 Hz, 1H), 7.24 (dd, J = 1.6, 8.0 Hz, 1H), 7.43 (dd, J = 1.2, 10.4 Hz,1H), 7.59-7.66 (m, 2H), 7.84 (d, J = 2.4 Hz, 1H). 62

417 ¹H N MR (400 MHz, CD₃OD): δ 2.27-2.31 (m, 2H), 3.44-.347 (m, 2H),3.60-3.64 (m, 5H), 3.70- 3.73 (m, 2H), 3.91-3.94 (m, 5H), 6.81 (d, J =8.4 Hz, 1H), 7.53 (d, J = 8.4 Hz, 2H), 7.58 (dd, J = 2.4, 8.8 Hz, 1H),7.64 (d, J = 8.8 Hz, 2H), 7.84 (d, J = 1.2 Hz, 1H). 63

448 ¹H NMR (400 MHz, CD₃OD): δ 2.31-2.33 (m, 2H), 3.27 (s, 6H) 3.44-.347(m, 2H), 3.60-3.67 (m, 5H), 3.73-3.76 (m, 2H), 3.96-3.99 (m, 2H), 6.81(d, J = 8.4 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.58 (dd, J = 2.4, 8.8Hz, 1H), 7.64 (d, J = 8.8 Hz, 2H), 7.84 (d, J = 1.2 Hz, 1H). 64

406 ¹H NMR (400 MHz, CD₃OD): δ 3.39 (s, 3H), 3.67 (s, 3H), 4.09- 4.10(m, 1H), 4.17-4.21 (m, 2H), 4.55-4.59 (m, 2H), 6.74 (d, J = 8.8 Hz, 2H),6.96 (d, J = 8.8 Hz, 2H), 7.10 (dd, J = 1.6, 8.4 Hz, 1H), 7.23 (dd, J =1.6, 10.8 Hz, 1H), 7.45 (dd, J = 6.8, 8.0 Hz, 1H). 65

472 ¹H NMR (400 MHz, CD₃OD): δ 1.85-1.98 (m, 2H), 2.23-2.27 (m, 2H),2.78 (s, 3H), 3.38-3.40 (m, 1H), 3.62 (s, 3H), 3.90-3.95 (m, 2H), 4.41(s, 3H), 7.26 (d, J = 10.8 Hz, 1H), 7.40 (d, J = 13.6 Hz, 1H), 7.49-7.57(m, 2H), 7.65 (dd, J = 6.8, 11.6 Hz, 1H), 7.73 (s, 1H), 8.66 (s, 1H). 66

458 ¹H NMR (400 MHz, CD₃OD): δ 2.19-2.20 (m, 2H), 3.33-3.36 (m, 2H),3.50-3.53 (m, 5H), 3.60- 3.63 (m, 2H), 3.83-3.85 (m, 2H), 4.42 (s, 3H),7.13 (d, J = 7.6 Hz, 1H), 7.28 (d, J = 10.4 Hz, 1H), 7.34 (d, J = 9.2Hz, 1H), 7.40 (t, J = 7.2 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.58 (s,1H), 8.48 (s, 1H). 67

430 ¹H NMR (400 MHz, CD₃OD): δ 2.33-2.35 (m, 2H), 3.27 (s, 6H),3.46-3.49 (m, 2H), 3.62-3.66 (m, 5H), 3.75-3.78 (m, 2H), 3.98-4.02 (m,2H), 6.73 (d, J = 9.2 Hz, 1H), 7.67-7.72 (m, 5H), 7.80 (s, 1H). 68

490 ¹H NMR (400 MHz, CD₃OD): δ 1.89-1.97 (m, 2H), 2.15-2.18 (m, 2H),3.15-3.21 (m, 2H), 3.43- 3.49 (m, 1H), 3.61 (s, 3H), 3.69- 3.71 (m, 4H),3.86-3.94 (m, 6H), 7.31 (d, J = 9.6 Hz, 1H), 7.41 (d, J = 8.0 Hz, 1H),7.59 (d, J = 10.0 Hz, 1H), 7.72-7.80 (m, 2H), 7.91 (s, 1H). 69

420 ¹H NMR (400 MHz, CD₃OD): δ 2.99-3.06 (m, 1H), 3.30-3.32 (m, 2H),3.49 (s, 3H), 3.78 (s, 3H), 4.10-4.15 (m, 2H), 4.46-4.52 (m, 2H), 6.83(d, J = 8.4 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 7.19 (dd, J = 2.0, 10.8Hz, 1H), 7.33 (dd, J = 2.0, 14.4 Hz, 1H), 7.45 (dd, J = 8.8, 10.8 Hz,1H). 70

434 ¹H NMR (400 MHz, CD₃OD): δ 2.78 (s, 3H), 3.07-3.10 (m, 1H),3.37-3.39 (m, 2H), 3.48 (s, 3H), 3.78 (s, 3H), 4.12-4.15 (m, 2H),4.47-4.52 (m, 2H), 6.84 (d, J = 8.8 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H),7.20 (d, J = 8.0 Hz, 1H), 7.33 (d, J = 10.4 Hz, 1H), 7.45 (t, J = 7.2Hz, 1H). 71

486 ¹H NMR (400 MHz, CD₃OD): δ 1.67-1.73 (m, 2H), 2.02-2.06 (m, 2H),2.33 (s, 6H), 2.41-2.45 (m, 1H), 2.95-3.03 (m, 2H), 3.59 (s, 3H),3.79-3.84 (m, 2H), 4.18 (s, 3H), 7.10 (dd, J = 2.0, 12.0 Hz, 1H), 7.23(dd, J = 1.6, 10.8 Hz, 1H), 7.37 (dd, J = 2.0, 14.4 Hz, 1H), 7.46-7.56(m, 3H), 8.11 (s, 1H). 72

486 ¹H NMR (400 MHz, DMSO-d₆): δ 1.50-1.63 (m, 2H), 1.85-1.89 (m, 2H),2.18 (s, 6H), 2.21-2.27 (m, 1H), 2.85-2.92 (m, 2H), 3.45 (s, 3H),3.67-3.71 (m, 2H), 4.00 (s, 3H), 7.09-7.17 (m, 2H), 7.39 (dd, J = 1.6,14.4 Hz, 1H), 7.51- 7.54 (m, 2H), 7.69 (t, J = 9.2 Hz, 1H), 7.97 (s,1H). 73

443 ¹H NMR (300 MHz, CD₃OD): δ 1.85-1.91 (m, 2H), 2.11-2.16 (m, 2H),3.06-3.14 (m, 2H), 3.36- 3.40 (m, 1H), 3.57 (s, 3H), 3.81- 3.85 (m, 2H),6.85 (d, J = 8.4 Hz, 2H), 7.22 (d, J = 3.0 Hz, 1H), 7.24 (s, 1H),7.33-7.48 (m, 4H). 74

456 ¹H NMR (400 MHz, CD₃OD): δ 1.53-1.56 (m, 2H), 1.88-1.91 (m, 2H),2.87-2.95 (m, 3H), 3.49 (s, 3H), 3.62 (d, J = 13.6 Hz, 2H), 3.69 (s,3H), 6.26 (s, 1H), 6.81 (d, J = 4.0 Hz, 1H), 7.04-7.35 (m, 6H). 75

442 ¹H NMR (400 MHz, CD₃OD): δ 1.88-1.94 (m, 2H), 2.13-2.16 (m, 2H),3.05-3.16 (m, 2H), 3.33- 3.42 (m, 1H), 3.60 (s, 3H), 3.84- 3.86 (m, 2H),6.74-6.77 (m, 1H), 7.20-7.50 (m, 7H). 76

457 ¹H NMR (400 MHz, CD₃OD): δ 1.51-1.54 (m, 2H), 1.88-1.91 (m, 2H),2.84-2.97 (m, 3H), 3.49 (s, 3H), 3.62-3.65 (m, 5H), 6.31 (d, J = 2.8 Hz,1H), 6.61 (d, J = 8.0 Hz, 1H), 7.06 (d, J = 3.2 Hz, 1H), 7.13-7.18 (m,3H), 7.27 (d, J = 10.8 Hz, 1H), 7.33-7.37 (m, 2H). 77

444 ¹H NMR (400 MHz, CD₃OD): δ 1.65-1.68 (m, 2H), 2.01-2.04 (m, 2H),2.98-3.12 (m, 3H), 3.63 (s, 3H), 3.78-3.82 (m, 2H), 6.94 (d, J = 8.4 Hz,1H), 7.24 (d, J = 8.0 Hz, 1H), 7.43-7.45 (m, 2H), 7.50 (t, J = 7.2 Hz,1H), 7.72 (d, J = 8.4 Hz, 1H), 8.06 (s, 1H). 78

452 ¹H NMR (300 MHz, CD₃OD): δ 1.84-1.91 (m, 1H), 2.01-2.06 (m, 1H),3.00-3.08 (m, 2H), 3.16- 3.21 (m, 1H), 3.58 (s, 3H), 3.75- 3.82 (m, 4H),3.93-4.01 (m, 1H), 4.70-4.82 (m, 1H), 6.86 (d, J = 9.0 Hz, 2H), 7.06 (d,J = 8.7 Hz, 2H), 7.24 (dd, J = 0.9, 8.1 Hz, 1H), 7.34 (dd, J = 1.5, 10.8Hz, 1H), 7.54-7.58 (m, 1H). 79

452 ¹H NMR (400 MHz, CD₃OD): δ 1.87-1.91 (m, 1H), 2.03-2.07 (m, 1H),3.02-3.08 (m, 2H), 3.19- 3.29 (m, 1H), 3.59 (s, 3H), 3.77- 3.83 (m, 4H),3.95-4.01 (m, 1H), 4.73-4.85 (m, 1H), 6.87 (d, J = 8.8 Hz, 2H), 7.07 (d,J = 8.8 Hz, 2H), 7.26 (dd, J = 1.2, 8.4 Hz, 1H), 7.36 (dd, J = 1.2, 10.8Hz, 1H), 7.56 (t, J = 6.8 Hz, 1H). 80

486 ¹H NMR (400 MHz, CD₃OD): δ 1.71-1.77 (m, 2H), 2.05-2.08 (m, 2H),2.38 (s, 6H), 2.45-2.48 (m, 1H), 2.98-3.05 (m, 2H), 3.61 (s, 3H),3.83-3.86 (m, 2H), 4.20 (s, 3H), 6.93 (dd, J = 1.2, 8.8 Hz, 1H), 7.27(dd, J = 1.2, 7.6 Hz, 1H), 7.36-7.41 (m, 2H), 7.49-7.53 (m, 1H), 7.66(d, J = 8.8 Hz, 1H), 8.18 (s, 1H). 81

477 ¹H NMR (400 MHz, CD₃OD): δ 2.03-2.06 (m, 2H), 2.25-2.27 (m, 2H),2.98 (s, 6H), 3.14-3.20 (m, 2H), 3.36 (s, 6H), 3.56-3.60 (m, 1H), 3.62(s, 3H), 4.01-4.04 (m, 2H), 7.49 (d, J = 4.4 Hz, 1H), 7.67 (d, J = 10.0Hz, 1H), 7.75- 7.78 (m, 1H), 8.43 (s, 2H). 82

447 ¹H NMR (400 MHz, CD₃OD): δ 1.56-1.62 (m, 2H), 1.91-1.94 (m, 2H),2.25 (s, 6H), 2.31-2.37 (m, 1H), 2.41 (s, 3H), 2.87-2.93 (m, 2H), 3.47(s, 3H), 3.72-3.76 (m, 2H), 7.10 (dd, J = 1.2, 8.0 Hz, 1H), 7.15 (d, J =8.0 Hz, 1H), 7.28-7.31 (m, 1H), 7.49-7.52 (m, 2H), 7.80 (d, J = 2.0 Hz,1H). 83

462 ¹H NMR (400 MHz, CD₃OD): δ 1.92-2.04 (m, 2H), 2.24-2.26 (m, 2H),2.79 (s, 3H), 3.14-3.20 (m, 2H), 3.30 (s, 6H), 3.37-3.40 (m, 1H), 3.61(s, 3H), 3.94-3.98 (m, 2H), 7.16 (d, J = 9.6 Hz, 1H), 7.42 (d, J = 8.0Hz, 1H), 7.61 (d, J = 10.0 Hz, 1H), 7.71-7.76 (m, 2H), 7.84 (d, J = 1.2Hz, 1H). 84

472 ¹H NMR (400 MHz, CDCl₃): δ 1.61-1.69 (m, 2H), 1.98-2.01 (m, 2H),2.32 (s, 6H), 2.32-2.33 (m, 1H), 2.88-2.94 (m, 2H), 3.53 (s, 3H),3.65-3.69 (m, 2H), 6.73 (dd, J = 1.2, 8.8 Hz, 1H), 7.00 (dd, J = 1.2,8.0 Hz, 1H), 7.19- 7.30 (m, 2H), 7.37 (s, 1H), 7.52 (d, J = 8.4 Hz, 1H),7.90 (s, 1H), 10.65 (br, 1H). 85

455 ¹H NMR (400 MHz, CD₃OD): δ 1.88-1.92 (m, 2H), 2.15-2.18 (m, 2H),3.12-3.18 (m, 2H), 3.40- 3.46 (m, 1H), 3.85-3.88 (m, 5H), 6.81 (d, J =8.4 Hz, 1H), 6.98- 7.05 (m, 2 H), 7.25 (dd, J = 1.2, 8.4 Hz, 1H), 7.42(d, J = 11.2 Hz, 1H), 7.61 (t, J = 7.2 Hz, 1H). 86

455 ¹H NMR (400 MHz, CD₃OD): δ 1.88-1.92 (m, 2H), 2.14-2.17 (m, 2H),3.11-3.17 (m, 2H), 3.42- 3.47 (m, 1H), 3.59 (s, 3H), 3.85- 3.88 (m, 2H),6.81 (d, J = 8.4 Hz, 1H), 6.98-7.05 (m, 2 H), 7.25 (dd, J = 1.2, 8.0 Hz,1H), 7.42 (d, J = 10.4 Hz, 1H), 7.60 (dd, J = 0.8, 7.6 Hz, 1H). 87

472 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.79 (br. s., 2 H) 2.11 (br. s., 2H) 2.97 (br. s., 2 H) 3.10- 3.31 (m, 1 H) 3.46 (br. s., 3 H) 3.74 (d, J= 18.19 Hz, 2 H) 4.03 (br. s., 3 H) 7.12 (d, J = 13.39 Hz, 1 H)7.40-7.61 (m, 4 H) 7.71 (br. s., 1 H) 7.87-8.07 (m, 1 H) 9.15 (br. s., 2H). 88

444 ¹H NMR (400 MHz, Methanol- d₄): δ ppm 1.84 (d, J = 13.39 Hz, 2 H)2.11 (d, J = 13.14 Hz, 2 H) 3.05-3.17 (m, 2 H) 3.35-3.40 (m, 1 H) 3.59(s, 3 H) 3.83 (d, J = 14.40 Hz, 2 H) 7.17 (d, J = 8.08 Hz, 2 H) 7.41 (d,J = 10.61 Hz, 1 H) 7.44-7.52 (m, 2 H) 7.57 (s, 1 H) 7.98 (s, 1 H) 8.54(br. s., 1 H). 89

419 ¹H NMR (400 MHz, Methanol- d₄): δ ppm 1.74-1.96 (m, 2 H) 2.11 (d, J= 12.13 Hz, 2 H) 3.08 (q, J = 11.54 Hz, 2 H) 3.38 (br. s., 1 H) 3.57(br. s., 3 H) 3.71- 3.93 (m, 2 H) 6.64 (d, J = 8.08 Hz, 2 H) 6.86 (d, J= 8.08 Hz, 2 H) 7.07-7.17 (m, 1 H) 7.18- 7.30 (m, 1 H) 7.31-7.43 (m, 1H). 90

433 ¹H NMR (400 MHz, Methanol- d₄): δ ppm 1.72-1.93 (m, 2 H) 2.09 (d, J= 11.62 Hz, 2 H) 2.75 (s, 3 H) 2.99-3.14 (m, 2 H) 3.36- 3.43 (m, 1 H)3.56 (s, 3 H) 3.78 (d, J = 12.38 Hz, 2 H) 6.54 (d, J = 7.83 Hz, 2 H)6.89 (d, J = 7.83 Hz, 2 H) 7.27 (d, J = 8.34 Hz, 1 H) 7.32-7.43 (m, 1 H)7.48- 7.62 (m, 1 H). 91

451 ¹H NMR (400 MHz, Methanol- d₄): δ ppm 1.86 (d, J = 11.87 Hz, 2 H)2.12 (d, J = 11.12 Hz, 2 H) 2.96 (s, 3 H) 3.11 (t, J = 12.25 Hz, 2 H)3.40 (br. s., 1 H) 3.57 (s, 3 H) 3.84 (d, J = 12.38 Hz, 2 H) 6.90 (d, J= 8.59 Hz, 1 H) 7.05 (t, J = 8.46 Hz, 1 H) 7.12 (d, J = 12.38 Hz, 1 H)7.26 (d, J = 8.34 Hz, 1 H) 7.40 (d, J = 10.61 Hz, 1 H) 7.60 (t, J = 7.20Hz, 1 H). 92

463 ¹H NMR (400 MHz, Chloroform-d): δ ppm 1.71 (m, J = 11.37 Hz, 2 H)1.74 (br. s., 1 H) 2.04 (d, J = 11.87 Hz, 2 H) 2.38 (br. s., 6 H) 2.96(t, J = 12.76 Hz, 2 H) 3.55 (s, 3 H) 3.71 (d, J = 12.88 Hz, 2 H) 3.91(s, 3 H) 6.73 (d, J = 8.59 Hz, 1 H) 7.12 (d, J = 7.83 Hz, 1 H) 7.34 (d,J = 10.11 Hz, 1 H) 7.43 (t, J = 7.07 Hz, 1 H) 7.53 (d, J = 8.34 Hz, 1 H)7.81 (br. s., 1 H). 93

467 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.32 (td, J = 7.01, 1.39 Hz, 3 H)1.58 (d, J = 11.62 Hz, 2 H) 1.92 (d, J = 11.62 Hz, 2 H) 2.80 (s, 3 H)2.91-3.03 (m, 2 H) 3.08 (br. s., 1 H) 3.69 (d, J = 10.36 Hz, 2 H)4.29-4.40 (m, 2 H) 6.86 (s, 1 H) 6.89 (d, J = 8.08 Hz, 1 H) 7.21 (d, J =8.08 Hz, 1 H) 7.56 (d, J = 1.77 Hz, 1 H) 7.81-7.86 (m, 1 H) 8.33 (s, 3H). 94

449 ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.28 (td, J = 7.01, 2.40 Hz, 3 H)1.58 (br. s., 2 H) 1.89 (br. s., 2 H) 2.92-3.02 (m, 2 H) 3.07 (br. s., 1H) 3.43 (s, 3 H) 3.68 (d, J = 13.39 Hz, 2 H) 4.21- 4.29 (m, 2 H) 6.73(d, J = 3.79 Hz, 1 H) 6.80 (s, 1 H) 6.93 (d, J = 7.83 Hz, 1 H) 7.20 (d,J = 8.59 Hz, 1 H) 7.54 (d, J = 8.08 Hz, 1 H) 7.80-7.85 (m, 1 H) 8.31 (s,3 H). 95

448 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.31 (t, J = 6.69 Hz, 3 H) 1.73 (d,J = 9.09 Hz, 2 H) 2.00 (d, J = 12.13 Hz, 2 H) 2.99 (t, J = 12.51 Hz, 2H) 3.28 (br. s., 1 H) 3.43 (s, 3 H) 3.71 (d, J = 12.38 Hz, 2 H)3.95-4.06 (m, 2 H) 6.83 (d, J = 8.08 Hz, 2 H) 7.01 (d, J = 8.59 Hz, 2 H)7.18 (d, J = 8.59 Hz, 1 H) 7.41 (d, J = 10.86 Hz, 1 H) 7.61 (m, 1 H)7.79 (t, J = 7.83 Hz, 1 H) 8.07 (br. s., 3 H). 96

464 ¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.74 (d, J = 10.36 Hz, 2 H) 2.00 (d,J = 11.62 Hz, 2 H) 2.99 (t, J = 12.25 Hz, 2 H) 3.43 (s, 3 H) 3.64 (br.s., 2 H) 3.71 (d, J = 11.87 Hz, 2 H) 4.07 (br. s., 2 H) 6.85 (d, J =8.34 Hz, 2 H) 7.01 (d, J = 8.34 Hz, 2 H) 7.18 (d, J = 8.08 Hz, 1 H) 7.41(d, J = 10.36 Hz, 1 H) 7.58-7.67 (m, 1 H) 7.79 (t, J = 7.45 Hz, 1 H)8.14 (br. s., 3 H). 97

446 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.87 (d, J = 11.12 Hz, 2 H)2.13 (d, J = 12.13 Hz, 2 H) 3.04-3.21 (m, 2 H) 3.38 (d, J = 10.61 Hz, 1H) 3.57 (s, 3 H) 3.77- 3.88 (m, 4 H) 4.02 (br. s., 2 H) 6.86 (d, J =7.83 Hz, 2 H) 7.04 (d, J = 8.34 Hz, 2 H) 7.47- 7.53 (m, 2 H) 7.57 (d, J= 7.58 Hz, 2 H). 98

478 ¹H NMR (400 MHz, DMSO-d₆) δ ppm: 1.74 (d, J = 11.87 Hz, 2 H) 2.00(d, J = 12.38 Hz, 2 H) 2.99 (t, J = 12.25 Hz, 2 H) 3.28 (br. s., 1 H)3.43 (s, 3 H) 3.44- 3.54 (m, 2 H) 3.70 (m, 5 H) 3.90- 4.05 (m, 2 H) 6.85(d, J = 8.34 Hz, 2 H) 7.01 (d, J = 7.83 Hz, 2 H) 7.18 (d, J = 8.08 Hz, 1H) 7.42 (d, J = 10.61 Hz, 1 H) 7.79 (t, J = 7.20 Hz, 1 H) 8.11 (br. s.,3 H). 99

448 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.73 (d, J = 11.37 Hz, 2 H) 2.00(d, J = 12.63 Hz, 2 H) 2.64- 2.76 (m, 2 H) 3.00 (t, J = 12.13 Hz, 2 H)3.29 (br. s., 1 H) 3.43 (br. s., 3 H) 3.48 (d, J = 9.85 Hz, 2 H)3.69-3.77 (m, 2 H) 7.00 (d, J = 7.33 Hz, 2 H) 7.13 (d, J = 7.83 Hz, 2 H)7.18 (d, J = 8.34 Hz, 1 H) 7.34-7.42 (m, 1 H) 7.75-7.80 (m, 1 H) 8.06(br. s., 3 H). 100

434 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.73 (d, J = 12.13 Hz, 2 H)1.94-2.03 (m, 2 H) 3.00 (br. s., 2 H) 3.29 (br. s., 1 H) 3.44 (s, 3 H)3.48 (d, J = 9.60 Hz, 2 H) 3.70 (br. s., 2 H) 7.06 (d, J = 7.33 Hz, 2 H)7.16-7.25 (m, 3 H) 7.41 (d, J = 10.86 Hz, 1 H) 7.75-7.82 (m, 1 H) 8.05(br. s., 3 H). 101

422 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.78-1.94 (m, 2 H) 2.13 (d, J =11.87 Hz, 2 H) 3.10 (t, J = 12.51 Hz, 2 H) 3.39 (d, J = 12.13 Hz, 1 H)3.57 (s, 3 H) 3.73-3.93 (m, 2 H) 6.99- 7.09 (m, 2 H) 7.12-7.25 (m, 3 H)7.37 (d, J = 10.36 Hz, 1 H) 7.57 (t, J = 6.95 Hz, 1 H). 102

422 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.79-1.93 (m, 2 H) 2.13 (d, J =12.38 Hz, 2 H) 3.11 (t, J = 12.63 Hz, 2 H) 3.39 (d, J = 11.62 Hz, 1 H)3.57 (s, 3 H) 3.85 (d, J = 13.64 Hz, 2 H) 6.89 (d, J = 7.83 Hz, 1 H)6.96-7.07 (m, 2 H) 7.23 (d, J = 8.34 Hz, 1 H) 7.25-7.33 (m, 1 H) 7.38(d, J = 10.36 Hz, 1 H) 7.58 (t, J = 7.20 Hz, 1 H). 103

440 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.77-1.95 (m, 2 H) 2.13 (d, J =11.62 Hz, 2 H) 3.12 (t, J = 12.76 Hz, 2 H) 3.36-3.45 (m, 1 H) 3.55 (s, 3H) 3.86 (d, J = 13.64 Hz, 2 H) 6.78 (d, J = 6.57 Hz, 2 H) 6.89 (t, J =9.22 Hz, 1 H) 7.24 (d, J = 8.08 Hz, 1 H) 7.43 (d, J = 10.11 Hz, 1 H)7.62 (t, J = 7.07 Hz, 1 H). 104

440 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.79-1.93 (m, 2 H) 2.12 (d, J =11.62 Hz, 2 H) 3.11 (t, J = 12.63 Hz, 2 H) 3.33-3.49 (m, 1 H) 3.57 (s, 3H) 3.85 (d, J = 13.64 Hz, 2 H) 6.87 (br. s., 1 H) 7.11-7.25 (m, 3 H)7.42 (d, J = 10.36 Hz, 1 H) 7.60 (t, J = 7.20 Hz, 1 H). 105

482 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.79-1.92 (m, 2 H) 2.12 (d, J =11.62 Hz, 2 H) 3.05-3.29 (m, 5 H) 3.40 (br. s., 1 H) 3.58 (s, 3 H) 3.80-3.94 (m, 2 H) 7.16 (d, J = 7.58 Hz, 1 H) 7.36-7.48 (m, 3 H) 7.58 (t, J =7.20 Hz, 1 H) 7.87 (d, J = 8.08 Hz, 2 H). 106

438 ¹H NMR (400 MHz, Chloroform-d): δ ppm 1.89 (d, J = 11.12 Hz, 2 H)2.16 (d, J = 10.86 Hz, 2 H) 3.05 (t, J = 11.87 Hz, 2 H) 3.28 (br. s., 1H) 3.55 (s, 3 H) 3.71 (d, J = 12.13 Hz, 2 H) 7.04 (d, J = 8.34 Hz, 1 H)7.10 (d, J = 8.08 Hz, 2 H) 7.27-7.30 (m, 1 H) 7.33-7.44 (m, 2 H) 8.31(br. s., 1 H). 107

448 ¹H NMR (400 MHz, Methanol-d₄): δ ppm 1.81-1.94 (m, 2 H) 2.13 (d, J =12.13 Hz, 2 H) 3.12 (t, J = 12.38 Hz, 2 H) 3.36 (s, 3 H) 3.41 (br. s., 1H) 3.58 (s, 3 H) 3.84 (d, J = 12.63 Hz, 2 H) 4.45 (s, 2 H) 7.14 (d, J =7.58 Hz, 2 H) 7.23 (d, J = 7.83 Hz, 1 H) 7.28 (d, J = 7.83 Hz, 2 H) 7.34(d, J = 10.61 Hz, 1 H) 7.55 (t, J = 7.20 Hz, 1 H). 108

328 ¹H NMR (400 MHz, DMSO-d₆): δ ppm 1.62-1.78 (m, 2 H) 2.00 (d, J =11.87 Hz, 2 H) 3.02 (t, J = 12.00 Hz, 2 H) 3.32 (s, 3 H) 3.76 (d, J =12.88 Hz, 2 H) 6.87 (s, 1 H) 7.95 (br. s., 3 H) 8.01- 8.08 (m, 1 H)8.08-8.12 (m, 1 H) 8.16 (d, J = 11.12 Hz, 1 H). 109

368 ¹HNMR (400 MHz, CD₃OD): δ 0.37-0.39 (m, 2H), 0.60-0.65 (m, 2H),1.29-1.32 (m, 1H), 1.59-1.64 (m, 2H), 2.10-2.14 (m, 2H), 3.07-3.14 (m,2H), 3.43-3.47 (m, 1H), 6.81 (d, J = 7.2 Hz, 2H), 4.92-4.95 (m, 2 H),6.66 (s, 1H), 7.84-7.88 (m, 1H), 7.99-8.05 (m, 2H). 110

352 ¹H NMR (400 MHz, CD₃OD): δ 1.40-1.41 (m, 2H), 1.81-1.84 (m, 2H),2.75-2.78 (m, 1H), 2.89-2.95 (m, 2H), 3.37 (s, 3H), 3.65-3.68 (m, 2H),3.77 (s, 1H), 7.66 (t, J = 8.0 Hz, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.96(d, J = 8.4 Hz, 1H). 111

442 ¹H NMR (400 MHz, DMSO-d₆): 1.75-1.83 (m, 2H), 2.06 (d, J = 10.8 Hz,2H), 2.99 (t, J = 11.6 Hz, 2H), 3.28-3.30 (m, 1H), 3.42 (s, 3H),3.68-3.74 (m, 5H), 6.85 (d, J = 8.0 Hz, 2H), 7.02- 7.08 (m, 3H),7.28-7.45 (m, 2H), 8.38-8.44 (m, 2H). 112

429 ¹H NMR (400 MHz, CD₃OD): 1.09-1.17 (m, 2H), 1.57-1.62 (m, 2H),2.46-2.56 (m, 3H), 2.96-3.03 (m, 2H), 3.35 (s, 3H), 3.78 (s, 3H), 6.02(s, 1H), 6.37 (s, 1H), 6.79-6.97 (m, 3H), 7.13-7.27 (m, 2H), 7.42-7.52(m, 2H). 113

431 ¹H NMR (400 MHz, CD₃OD): 1.06-1.17 (m, 2H), 1.57-1.62 (m, 2H),2.49-2.56 (m, 3H), 2.96-3.09 (m, 2H), 3.36 (s, 3H), 3.78 (s, 3H), 6.07(s, 1H), 6.40 (s, 1H), 6.97-7.20 (m, 4H), 7.27 (d, J = 11.8 Hz, 1H),7.45 (s, 1H), 7.61-7.66 (m, 1H). 114

414 ¹H NMR (400 MHz, CD₃OD): δ 152-1.55 (m, 2H), 1.87-1.90 (m, 2H),2.83-3.95 (m, 3H), 3.49 (s, 3H), 3.59-3.67 (m, 5H), 6.23 (s, 1H), 6.81(d, J = 8.4 Hz, 1H), 6.98-7.08 (m, 4H), 8.14 (t, J = 8.0 Hz, 1H), 7.24(m, 3H). 115

410 ¹H NMR (400 MHz, CD₃OD): δ 1.90-2.05 (m, 2H), 2.16-2.19 (m, 2H),3.12-3.20 (m, 2H), 3.44-3.49 (m, 1H), 3.62 (s, 3H), 3.88 (s, 3H),3.90-3.92 (m, 2H), 6.88-6.90 (m, 1H), 7.02-7.06 (m, 1H), 7.08-7.13 (m,1H), 8.07 (d, J = 6.0 Hz, 2H), 8.79 (d, J = 6.0 Hz, 2H). 116

415 ¹H NMR (400 MHz, CD₃OD): δ 1.50-1.53 (m, 2H), 1.86-1.89 (m, 2H),2.82-3.95 (m, 3H), 3.48 (s, 3H), 3.60-3.69 (m, 5H), 6.24 (s, 1H), 6.81(d, J = 8.4 Hz, 1H), 7.03 (s, 1H), 7.18-7.24 (m 4H), 8.17 (t, J = 4.4Hz, 1H). 117

444 ¹H NMR (400 MHz, CD₃OD): δ 1.62-1.68 (m, 2H), 2.01-2.03 (m, 2H),2.96-3.06 (m, 3H), 3.55 (s, 3H), 3.71-3.74 (m, 5H), 3.81 (s, 3H), 6.36(d, J = 3.2 Hz, 1H), 6.65 (d, J = 8.8 Hz, 1H), 6.93 (t, J = 8.8 Hz, 1H),7.13 (d, J = 3.2 Hz, 1H),7.29- 7.38 (m, 4H). 118

434 ¹H NMR (300 MHz, CD₃OD): δ 1.84-1.89 (m, 2H), 2.12-2.16 (m, 2H),3.13 (t, J = 12.0 Hz, 2H), 3.31-3.41 (m, 1H), 3.57 (s, 3H), 3.84 (s,3H), 3.84-3.86 (m, 2H), 6.79-6.82 (m, 1H), 6.93-7.00 (m, 2 H), 7.38 (t,J = 7.5 Hz, 1H), 7.54 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 7.8 Hz, 1H),7.78 (s, 1H). 119

434 ¹H NMR (300 MHz, DMSO-d₆): δ 1.70-1.74 (m, 2H), 1.99-2.03 (m, 2H),2.95 (t, J = 12.0 Hz, 2H), 3.23-3.24 (m, 1H), 3.44 (s, 3H), 3.74 (s,3H), 3.84-3.86 (m, 2H), 6.68-6.70 (m, 1H), 6.91-6.96 (m, 2 H), 7.31-7.34(m, 1H), 7.40-7.59 (m, 2H), 7.77-7.80 (m, 1H), 8.34 (m, 3H).

Preparation 120A:[1-(5-chloro-4-cyano-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl)-piperidin-4-yl]-carbamicacid tert-butyl ester

A mixture ofN-[1-(5,6-dichloro-3-methyl-4-oxo(3-hydropyrimidin-2-yl))(4-piperidyl)](tert-butoxy)carboxamide(2.4 g, 6.38 mmol), Zn(CN)₂ (388 mg, 3.32 mmol) and Pd(PPh₃)₄ (740 mg,0.64 mmol) in DMF (20 mL) was stirred at 130° C. for 5 h under N₂atmosphere. The reaction mixture was cooled to RT and filtered. Thefiltrate was concentrated in vacuo, and the residue was purified bypreparative HPLC to give 200 mg of the title product (9%). [M+H] Calc'dfor C₁₆H₂₂C₁N₅O₃, 368; Found, 368.

Preparation 120B:{1-[4-cyano-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of[1-(5-chloro-4-cyano-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl)-piperidin-4-yl]-carbamicacid tert-butyl ester (200 mg, 0.54 mmol),3-fluoro-4-methoxybenzeneboronic acid (278 mg, 1.63 mmol), Pd(dppf)₂Cl₂(119 mg, 0.16 mmol), and Na₂CO₃ (173 mg, 1.63 mmol) in dioxane (5 mL)and H₂O (1 mL) was degassed with N₂ and stirred at 145° C. in themicrowave for 2 h. The reaction mixture was cooled to RT and filtered.The filtrate was concentrated in vacuo and the residue purified bypreparative HPLC to give 110 mg of the desired product (45%). [M+H]Calc'd for C₂₃H₂₈FN₅O₄, 458; Found, 458.

Example 120:2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidine-4-carbonitrile

A mixture of{1-[4-cyano-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (100 mg, 0.23 mmol) in EA (5 mL) was added a 5NHCl solution in EA (5 mL) was stirred at RT for 2 h. The solvent wasconcentrated in vacuo to give 85 mg of the title product as the HCl salt(93%). ¹H NMR (400 MHz, CD₃OD): δ 1.71-1.75 (m, 2H), 1.89-2.03 (m, 2H),2.96-3.02 (m, 2H), 3.27-3.31 (m, 1H), 3.42 (s, 3H), 3.69-3.73 (m, 2H),3.83 (s, 3H), 7.06 (t, J=8.0 Hz, 1H), 7.17-2.01 (m, 2H). [M+H] Calc'dfor C₁₈H₂₀FN₅O₂, 358; Found, 358.

Preparation 121A:{1-[5-cyano-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of{1-[5-chloro-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (460 mg, 1 mmol), Zn(CN)₂ (175 mg, 1.5 mmol) andPd(PPh₃)₄ (116 mg, 0.0.1 mmol) in DMF (5 mL) was stirred 4 h at 150° C.under N₂ atmosphere. The mixture was cooled to RT and filtered. Thefiltrate was concentrated in vacuo, and the residue purified bypreparative HPLC to give 150 mg of the title product as a yellow solid(33%). [M+H] Calc'd for C₂₃H₂₅FN₆O₃, 453; Found, 453.

Example 121:2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidine-5-carbonitrile

To a mixture of{1-[5-cyano-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5 NHCl solution in EA (5 mL), and the mixture was stirred at RT for 2 h.The solvent was concentrated in vacuo to give 120 mg the title productas HCl salt (94%). ¹H NMR (400 MHz, CD₃OD): δ 1.67-1.72 (m, 2H),2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H),3.98-4.02 (m, 2H), 7.82-7.90 (m, 3H). [M+H] Calc'd for C₁₈H₁₇FN₆O, 353;Found, 353.

Preparation 122A: 4-cyano-3-fluoro-benzoyl chloride

A mixture of 4-cyano-3-fluoro-benzoic acid (2.0 g, 12.12 mmol) in SOCl₂(20 mL) was refluxed for 2 h, and SOCl₂ was removed in vacuo to give4-cyano-3-fluoro-benzoyl chloride (2.2 g, 99%). The crude was carried tothe next step without further purification.

Preparation 122B:3-(4-cyano-3-fluoro-phenyl)-2-(4-methoxy-phenyl)-3-oxo-propionic acidmethyl ester

To a solution of (4-methoxy-phenyl)-acetic acid (2.18 g, 12.12 mmol) inTHF (20 mL) was added LiHMDS (18.2 mL, 18.18 mmol) at −78° C. and themixture was stirred for 30 min. A solution of 4-cyano-3-fluoro-benzoylchloride (2.2 g, 12 mmol) in THF was added dropwise at −78° C.; and thereaction mixture was allowed to warm up to RT and stirred at overnight.Aqueous NH₄Cl was added and the aqueous was extracted with EA (3×). Thecombined organics were concentrated in vacuo and the residue waspurified by silica column chromatography (1:5, EA: PE) to give 1.8 g(45%) of the title compound. [M+H] Calc'd for C₁₈H₁₄FNO₄, 328; Found,328.

Preparation 122C:{1-[4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

A mixture of3-(4-cyano-3-fluoro-phenyl)-2-(4-methoxy-phenyl)-3-oxo-propionic acidmethyl ester (1.8 g, 5.5 mmol),(1-carbamimidoyl-piperidin-4-yl)-carbamic acid tert-butyl ester (2.6 g,9.2 mmol), DIEA (2.4 g, 18.3 mmol) in toluene (50 mL) was refluxedovernight. The solvent was concentrated in vacuo. The residue wassuspended in MeOH and the solids were filtered to give 100 mg (4%) ofthe title compound. [M+H] Calc'd for C₂₈H₃₀FN₅O₄, 520; Found, 520.

Example 122:4-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile

To a solution of{1-[4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-phenyl)-6-oxo-1,6-dihydro-pyrimidin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (50 mg, 0.096 mmol) in EA (10 mL) was added a 5MHCl solution in EA and the mixture was stirred at RT for 2 h. Thesolvent was removed in vacuo and the residue was purified by preparativeHPLC to give 18 mg (40%) of the title compound as the hydrochloridesalt. ¹H NMR (400 MHz, CD₃OD): δ 1.81-1.87 (m, 2H), 2.22-2.25 (m, 2H),3.34-3.38 (m, 2H), 3.56-3.60 (m, 1H), 3.78 (s, 3H), 4.61-4.64 (m, 2H),6.86 (d, J=7.2 Hz, 2H), 7.08 (d, J=8.4 Hz, 2H), 7.37-7.38 (m, 1H),7.51-7.53 (m, 1H), 7.74 (s, 1H). [M+H] Calc'd for C₂₃H₂₂FN₅O₂, 420;Found, 420.

II. Biological Evaluation Example 1a: In Vitro Enzyme InhibitionAssay—LSD-1

This assay determines the ability of a test compound to inhibit LSD1demethylase activity. E. coli expressed full-length human LSD1(Accession number 060341) was purchased from Active Motif (Cat#31334).

The enzymatic assay of LSD1 activity is based on TimeResolved-Fluorescence Resonance Energy Transfer (TR-FRET) detection. Theinhibitory properties of compounds to LSD1 were determined in 384-wellplate format under the following reaction conditions: 0.1-0.5 nM LSD1,50 nM H3K4me1-biotin labeled peptide (Anaspec cat #64355), 2 μM FAD inassay buffer of 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 0.5 mMTCEP, 0.2 mg/ml BSA. Reaction product was determined quantitatively byTR-FRET after the addition of detection reagent PhycolinkStreptavidin-allophycocyanin (Prozyme) and Europium-anti-unmodifiedhistone H3 lysine 4 (H3K4) antibody (PerkinElmer) in the presence ofLSD1 inhibitor such as 1.8 mM of Tranylcypromine hydrochloride (2-PCPA)in LANCE detection buffer (PerkinElmer) to final concentration of 12.5nM and 0.25 nM respectively.

The assay reaction was performed according to the following procedure: 2μL of the mixture of 150 nM H3K4me1-biotin labeled peptide with 2 μL of11-point serial diluted test compound in 3% DMSO were added to each wellof plate, followed by the addition of 2 μL of 0.3 nM LSD1 and 6 μM ofFAD to initiate the reaction. The reaction mixture was then incubated atroom temperature for one hour, and terminated by the addition of 6 μL of1.8 mM 2-PCPA in LANCE detection buffer containing 25 nM PhycolinkStreptavidin-allophycocyanin and 0.5 nM Europium-anti-unmodified H3K4antibody. Enzymatic reaction is terminated within 15 minutes if 0.5 LSD1enzyme is used in the plate. Plates were read by EnVision MultilabelReader in TR-FRET mode (excitation at 320 nm, emission at 615 nm and 665nm) after 1 hour incubation at room temperature. A ratio was calculated(665/615) for each well and fitted to determine inhibition constant(IC₅₀).

The ability of the compounds disclosed herein to inhibit LSD1 activitywas quantified and the respective IC₅₀ value was determined. Table 4provides the IC₅₀ values of various substituted heterocyclic compoundsdisclosed herein.

TABLE 4 Chemical Synthesis LSD1 Example Name IC₅₀ (μM) 14-(2-(4-aminopiperidin-1-yl)-1-methyl-6-oxo-5-p-tolyl-1,6-dihydropyrimidin-A 4-yl)benzonitrile 24-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-benzonitrile 34-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-benzonitrile 44-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)-6-oxo-1,6-A dihydro-pyrimidin-4-yl]-benzonitrile 54-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-benzonitrile 64-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 74-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-A oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 84-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 94-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 104-[2-(4-amino-piperidin-1-yl)-5-(6-ethyl-pyridin-3-yl)-1-methyl-6-oxo-1,6-A dihydro-pyrimidin-4-yl]-benzonitrile 112-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(4-methylamino-piperidin-1-A yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 122-fluoro-4-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 134-[2-(4-amino-piperidin-1-yl)-1-ethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-B fluoro-benzonitrile 144-[2-(4-amino-piperidin-1-yl)-5-cyclopentylethynyl-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 15[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy- Aphenyl)-6-oxo-6H-pyrimidin-1-yl]-acetic acid 162-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5-(4-methoxy-A phenyl)-6-oxo-6H-pyrimidin-1-yl]-acetamide 174-[2-(4-amino-piperidin-1-yl)-1-(3-hydroxy-propyl)-6-oxo-1,6-dihydro- Bpyrimidin-4-yl]-2-fluoro-benzonitrile 184-[2-(4-amino-piperidin-1-yl)-5-benzofuran-5-yl-1-methyl-6-oxo-1,6-dihydro-A pyrimidin-4-yl]-2-fluoro-benzonitrile 192-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-A dihydro-pyrimidine-5-carbonitrile 204-[2-(4-aminopiperidin-1-yl)-5-chloro-1-methyl-6-oxopyrimidin-4-yl]-2- Afluorobenzonitrile 212-fluoro-4-[1-methyl-2-(4-methylamino-piperidin-1-yl)-5-(6-methyl-pyridin-A 3-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 224-[2-(2,8-diaza-spiro[4.5]dec-8-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 234-{2-(4-aminopiperidyl)-1-methyl-6-oxo-5-[6-(trifluoromethyl)(3-pyridyl)] A hydropyrimidin-4-yl}-2-fluorobenzenecarbonitrile 244-[2-(4-aminopiperidyl)-1-methyl-5-(2-methyl(2H-indazol-5-yl))-6- Aoxohydropyrimidin-4-yl]benzenecarbonitrile 254-[2-((3R)-3-aminopiperidyl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6- Aoxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 264-[2-(4-aminopiperidyl)-5-(5-fluoro-6-methoxy(3-5,6-dihydropyridyl))-1-A methyl-6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 274-[2-((3R)-3-aminopyrrolidinyl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6-A oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 284-[2-((3S)-3-amino-piperidin-1-yl)-5- A(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 294-[2-((3S)-3-amino-pyrrolidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 304-[2-((3R)-3-aminopiperidyl)-5-(4-methoxyphenyl)-1-methyl-6-oxohydro- Apyrimidin-4-yl]-2-fluorobenzenecarbonitrile 314-[2-((3S)-3-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 324-[2-(4-amino-4-methyl-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-A methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 334-[2-(4-aminopiperidyl)-1-methyl-5-(1-methyl(1H-indazol-5-yl))-6- Aoxohydropyrimidin-4-yl]benzenecarbonitrile 344-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-2-fluoro-benzonitrile 354-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indazol-5-yl)-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 364-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 374-[2-(4-aminopiperidyl)-1-methyl-5-(2-methyl(2H-indazol-5-yl))-6- Aoxohydropyrimidin-4-yl]-2-fruorobenzenecarbonitrile 384-[2-(4-aminopiperidyl)-5-(3,5-difluoro-4-methoxyphenyl)-1-methyl-6- Aoxohydropyrimidin-4-yl]benzenecarbonitrile 394-[2-(4-aminopiperidyl)-6-(4-cyano-3-fluorophenyl)-3-methyl-4-oxo-3- Bhydropyrimidin-5-yl]benzoic acid 40{4-[2-(4-aminopiperidyl)-6-(4-cyanophenyl)-3-methyl-4-oxo(3-hydro Apyrimidin-5-yl)]-2-fluorophenyl}-N-methylcarboxamide 414-[2-(4-aminopiperidyl)-6-(4-cyanophenyl)-3-methyl-4-oxo(3-hydro Apyrimidin-5-yl)]-2-fluorobenzamide 424-[2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-(1-oxo-2,3-dihydro-1H- Aisoindol-5-yl)-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 433-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-C 1,6-dihydro-pyrimidin-5-yl]-benzoic acid 444-{5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3S)-(pyrrolidin-3- Aylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 454-{5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3R)-(pyrrolidin-3- Aylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 464-[2-[1,4]diazepan-1-yl-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-A dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 472-fluoro-4-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-piperazin-1-yl-A 1,6-dihydro-pyrimidin-4-yl]-benzonitrile 484-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-(piperidin-4-ylamino)-A 1,6-dihydro-pyrimidin-4-yl]-benzonitrile 494-[2-(4-amino-piperidin-1-yl)-2′-dimethylamino-1-methyl-6-oxo-1,6-dihydro-A [5,5′]bipyrimidinyl-4-yl]-2-fluoro-benzonitrile 505-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-5-yl]-pyridine-2-carboxylic acid methylamide 512-fluoro-4-{5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3S)-(pyrrolidin-3- Aylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 522-luoro-4-{5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3R)-(pyrrolidin-3- Aylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 532-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-(piperidin-4-yl Aamino)-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 542-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-(3S)-pyrrolidin- A3-ylmethyl-amino)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 552-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-piperidin-4-yl-amino)-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 562-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-pyrrolidin-3-ylmethyl-A amino)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 574-[2-(4-amino-piperidin-1-yl)-5-(6-dimethylamino-pyridin-3-yl)-1-methyl-6-A oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 582-fluoro-4-[5-(6-methoxy-pyridin-3-yl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 594-[2-(4-amino-piperidin-1-yl)-5-(4-dimethylamino-phenyl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 604-[2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-(6-pyrrolidin-1-yl-pyridin-3-A yl)-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 614-[2-[1,4]diazepan-1-yl-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 624-[2-[1,4]diazepan-1-yl-5-(6-methoxy-pyridin-3-yl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 634-[2-[1,4]diazepan-1-yl-5-(6-dimethylamino-pyridin-3-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 644-[2-(3-amino-azetidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 652-fluoro-4-[1-methyl-2-(4-methylamino-piperidin-1-yl)-5-(2-methyl-2H- Aindazol-5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 664-[2-[1,4]diazepan-1-yl-1-methyl-5-(2-methyl-2H-indazol-5-yl)-6-oxo-1,6-A dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 674-[2-[1,4]diazepan-1-yl-5-(6-dimethylamino-pyridin-3-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-benzonitrile 684-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-morpholin-4-yl-pyridin-3-yl)-6-A oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 694-[2-(3-aminomethyl-azetidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 702-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(3-methylaminomethyl- Aazetidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 714-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(2-methyl-2H-indazol-5-A yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 724-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indazol-5-A yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 734-[2-(4-amino-piperidin-1-yl)-5-(1H-indol-5-yl)-1-methyl-6-oxo-1,6-dihydro-A pyrimidin-4-yl]-2-fluoro-benzonitrile 744-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indol-5-yl)-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 754-[2-(4-amino-piperidin-1-yl)-5-(1H-indol-6-yl)-1-methyl-6-oxo-1,6-dihydro-A pyrimidin-4-yl]-2-fluoro-benzonitrile 764-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indol-6-yl)-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 774-[2-(4-amino-piperidin-1-yl)-5-(1H-indazol-6-yl)-1-methyl-6-oxo-1,6- Adihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 784-[2-((4R,3S)-4-amino-3-fluoro-piperidin-1-yl)-5-(4-methoxy-phenyl)-1- Amethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 794-[2-((4S,3R)-4-amino-3-fluoro-piperidin-1-yl)-5-(4-methoxy-phenyl)-1- Amethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 804-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(2-methyl-2H-indazol-6-A yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 814-[2′-dimethylamino-2-(4-dimethylamino-piperidin-1-yl)-1-methyl-6-oxo-1,6-A dihydro-[5,5′]bipyrimidinyl-4-yl]-2-fluoro-benzonitrile 824-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)-6-A oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 834-[5-(6-dimethylamino-pyridin-3-yl)-1-methyl-2-(4-methylamino-piperidin-1-A yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 844-[2-(4-dimethylamino-piperidin-1-yl)-5-(2H-indazol-6-yl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 854-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1- Adeuteratedmethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile864-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-deuteratedmethoxy-phenyl)-1-A methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 872-fluoro-4-[1-methyl-2-[4-(methylamino)piperidin-1-yl]-5- A(1-methylindazol-5-yl)-6-oxopyrimidin-4-yl]benzonitrile 884-[2-(4-aminopiperidin-1-yl)-5-(1H-indazol-5-yl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 894-[5-(4-aminophenyl)-2-(4-aminopiperidin-1-yl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 904-[2-(4-aminopiperidin-1-yl)-1-methyl-5-[4-(methylamino) Aphenyl]-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 914-[2-(4-aminopiperidin-1-yl)-5-[3-fluoro-4-(methylamino)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 924-[2-[4-(dimethylamino)piperidin-1-yl]-5-(6-methoxypyridin-3-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 934-[2-(4-aminopiperidin-1-yl)-5-(6-ethoxy-5-fluoropyridin-3-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 944-[2-(4-aminopiperidin-1-yl)-5-(6-ethoxypyridin-3-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 954-[2-(4-aminopiperidin-1-yl)-5-(4-ethoxyphenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 964-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethoxy)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 974-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethoxy)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]benzonitrile 984-[2-(4-aminopiperidin-1-yl)-5-[4-(2-methoxyethoxy)phenyl]-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 994-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethyl)phenyl]-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 1004-[2-(4-aminopiperidin-1-yl)-5-[4-(hydroxymethyl)phenyl]-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 1014-[2-(4-aminopiperidin-1-yl)-5-(4-fluorophenyl)-1-methyl-6-oxopyrimidin-4-A yl]-2-fluorobenzonitrile 1024-[2-(4-aminopiperidin-1-yl)-5-(3-fluorophenyl)-1-methyl-6-oxopyrimidin-4-A yl]-2-fluorobenzonitrile 1034-[2-(4-aminopiperidin-1-yl)-5-(3,5-difluorophenyl)-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 1044-[2-(4-aminopiperidin-1-yl)-5-(3,4-difluorophenyl)-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 1054-[2-(4-aminopiperidin-1-yl)-1-methyl-5-(4-methylsulfonylphenyl)- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1064-[2-(4-aminopiperidin-1-yl)-5-(4-chlorophenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1074-[2-(4-aminopiperidin-1-yl)-5-[4-(methoxymethyl)phenyl]-1-methyl-6- Aoxopyrimidin-4-yl]-2-fluorobenzonitrile 1084-[2-(4-aminopiperidin-1-yl)-1-methyl-6-oxopyrimidin-4-yl]-2- Afluorobenzonitrile 1094-[2-(4-amino-piperidin-1-yl)-1-cyclopropylmethyl-6-oxo-1,6-dihydro- Bpyrimidin-4-yl]-2-fluoro-benzonitrile 1104-[2-(4-amino-piperidin-1-yl)-1-cyclopropylmethyl-6-oxo-1,6-dihydro- Apyrimidin-4-yl]-2-fluoro-benzonitrile 1112-(4-amino-piperidin-1-yl)-6-(4-chloro-3-fluoro-phenyl)-5-(4-methoxy- Bphenyl)-3-methyl-3H-pyrimidin-4-one 1122-(4-amino-piperidin-1-yl)-6-(4-hydroxy-phenyl)-3-methyl-5-(1-methyl-1H-D indol-5-yl)-3H-pyrimidin-4-one 1132-(4-amino-piperidin-1-yl)-6-(4-fluoro-phenyl)-3-methyl-5-(1-methyl-1H-B indol-5-yl)-3H-pyrimidin-4-one 1142-(4-amino-piperidin-1-yl)-3-methyl-5-(1-methyl-1H-indol-5-yl)-6-phenyl-D 3H-pyrimidin-4-one 1152-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-3-methyl-6- Cpyridin-4-yl-3H-pyrimidin-4-one 1162-(4-amino-piperidin-1-yl)-3-methyl-5-(1-methyl-1H-indol-5-yl)-6-pyridin-4-B yl-3H-pyrimidin-4-one 1172-(4-amino-piperidin-1-yl)-6-(4-methoxy-phenyl)-3-methyl-5-(1-methyl-1H-C indol-5-yl)-3H-pyrimidin-4-one 1183-[2-(4-aminopiperidin-1-yl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6- Doxopyrimidin-4-yl]benzonitrile 1192-[2-(4-aminopiperidin-1-yl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl-6- Doxopyrimidin-4-yl]benzonitrile 1202-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-C 1,6-dihydro-pyrimidine-4-carbonitrile 1212-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-1,6-B dihydro-pyrimidine-5-carbonitrile 1224-[2-(4-aminopiperidin-1-yl)-5-(4-methoxyphenyl)-6-oxo-1H-pyrimidin-4- Ayl]-2-fluorobenzonitrile Note: Biochemical assay IC₅₀ data aredesignated within the following ranges: A: ≦0.10 μM; B: >0.10 μM to ≦1.0μM; C: >1.0 μM to ≦10 μM; D: >10 μM

Example 2: In Vitro Enzyme Inhibition Assay—MAO Selectivity

Human recombinant monoamine oxidase proteins MAO-A and MAO-B areobtained. MAOs catalyze the oxidative deamination of primary, secondaryand tertiary amines. In order to monitor MAO enzymatic activities and/ortheir inhibition rate by inhibitor(s) of interest, a fluorescent-based(inhibitor)-screening assay is performed.3-(2-Aminophenyl)-3-oxopropanamine (kynuramine dihydrobromide, SigmaAldrich), a non-fluorescent compound is chosen as a substrate.Kynuramine is a non-specific substrate for both MAOs activities. Whileundergoing oxidative deamination by MAO activities, kynuramine isconverted into 4-hydroxyquinoline (4-HQ), a resulting fluorescentproduct.

The monoamine oxidase activity was estimated by measuring the conversionof kynuramine into 4-hydroxyquinoline. Assays were conducted in 96-wellblack plates with clear bottom (Corning) in a final volume of 100 μl.The assay buffer was 100 mM HEPES, pH 7.5. Each experiment was performedin triplicate within the same experiment.

Briefly, a fixed amount of MAO (0.25 μg for MAO-A and 0.5 μg for AO-B)was incubated on ice for 15 minutes in the reaction buffer, in theabsence and/or in the presence of various concentrations of compounds asdisclosed herein (e.g., from 0 to 50 μM, depending on the inhibitorstrength). Tranylcypromine (Biomol International) was used as a controlfor inhibition.

After leaving the enzyme(s) interacting with the test compound, 60 to 90M of kynuramine was added to each reaction for MAO-B and MAO-A assayrespectively, and the reaction was left for 1 hour at 37° C. in thedark. The oxidative deamination of the substrate was stopped by adding50 μl of 2N NaOH. The conversion of kynuramine to 4-hydroxyquinoline wasmonitored by fluorescence (excitation at 320 nm, emission at 360 nm)using a microplate reader (Infinite 200, Tecan). Arbitrary units wereused to measure levels of fluorescence produced in the absence and/or inthe presence of test compound.

The maximum of oxidative deamination activity was obtained by measuringthe amount of 4-hydroxyquinoline formed from kynuramine deamination inthe absence of test compound and corrected for background fluorescence.The Ki (IC₅₀) of each inhibitor was determined at Vmax/2. Chemicalsynthesis examples 1-94, 101-106, 108-117, and 120-122 were tested inthe above described assay and found to have an IC₅₀ greater than 2micromolar.

Example 3: LSD1 CD11b Cellular Assay

To analyze LSD1 inhibitor efficacy in cells, a CD11b flow cytometryassay was performed. LSD1 inhibition induces CD11b expression in THP-1(AML) cells which is measured by flow cytometry. THP-1 cells were seededat 100,000 cells/well in 10% Fetal Bovine Serum containing RPMI 1640media in a 24 well plate with a final volume of 500 μL per well. LSD1test compounds were serially diluted in DMSO. The dilutions were addedto each well accordingly to a final concentration of 0.2% DMSO. Thecells were incubated at 37 degrees Celsius in 5% CO₂ for 4 days. 250 μLof each well was transferred to a well in a 96 well round bottom plate.The plate was centrifuged at 1200 rpm at 4 degrees Celsius in a BeckmanCoulter Alegra 6KR centrifuge for 5 minutes. The media was removedleaving the cells at the bottom of the wells. The cells were washed in100 μL cold HBSS (Hank's Balanced Salt Solution) plus 2% BSA (BovineSerum Albumin) solution and centrifuged at 1200 rpm at 4 degrees Celsiusfor 5 minutes. The wash was removed. The cells were resuspended in 100μL HBSS plus 2% BSA containing 1:15 dilution of APC conjugated mouseanti-CD11b antibody (BD Pharmingen Cat#555751) and incubated on ice for25 minutes. The cells were centrifuged and washed two times in 100 μlHBSS plus 2% BSA. After the final spin the cells were resuspended in 100μL HBSS plus 2% BSA containing 1 ug/mL DAPI(4′,6-diamidino-2-phenylindole). The cells were then analyzed by flowcytometry in a BD FACSAria machine. Cells were analyzed for CD11bexpression. The percent of CD11b expressing cells for each inhibitorconcentration was used to determine an IC₅₀ curve for each compoundanalyzed.

Table 5 provides the cellular IC₅₀ values of various substitutedheterocyclic compounds disclosed herein.

TABLE 5 Chemical Synthesis THP-1 Example Name IC₅₀ (μM) 14-(2-(4-aminopiperidin-1-yl)-1-methyl-6-oxo-5-p- Atolyl-1,6-dihydropyrimidin-4-yl)benzonitrile 24-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 34-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 44-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 54-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 64-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 74-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 84-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 94-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 104-[2-(4-amino-piperidin-1-yl)-5-(6-ethyl-pyridin-3-yl)-1-methyl- B6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 112-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 122-fluoro-4-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 144-[2-(4-amino-piperidin-1-yl)-5-cyclopentylethynyl-1-methyl-6-oxo- A1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 15[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5- C(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-acetic acid 162-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-5- A(4-methoxy-phenyl)-6-oxo-6H-pyrimidin-1-yl]-acetamide 184-[2-(4-amino-piperidin-1-yl)-5-benzofuran-5-yl-1-methyl-6-oxo- A1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 204-[2-(4-aminopiperidin-1-yl)-5-chloro-1-methyl- B6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 224-[2-(2,8-diaza-spiro[4.5]dec-8-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 234-{2-(4-aminopiperidyl)-1-methyl-6-oxo-5-[6-(trifluoromethyl)- A(3-pyridyl)] hydropyrimidin-4-yl}-2-fluorobenzenecarbonitrile 244-[2-(4-aminopiperidyl)-1-methyl-5-(2-methyl(2H-indazol-5-yl))- A6-oxohydropyrimidin-4-yl]benzenecarbonitrile 254-[2-((3R)-3-aminopiperidyl)-5-(3-fluoro-4-methoxyphenyl)- A1-methyl-6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 264-[2-(4-aminopiperidyl)-5-(5-fluoro-6-methoxy(3-5,6-dihydropyridyl))- A1-methyl-6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 274-[2-((3R)-3-aminopyrrolidinyl)-5-(3-fluoro-4-methoxyphenyl)-1-methyl- A6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 294-[2-((3S)-3-amino-pyrrolidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)- B1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 304-[2-((3R)-3-aminopiperidyl)-5-(4-methoxy-phenyl)-1-methyl- A 6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 314-[2-((3S)-3-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 324-[2-(4-amino-4-methyl-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 334-[2-(4-aminopiperidyl)-1-methyl-5-(1-methyl(1H-indazol-5-yl))- A6-oxohydropyrimidin-4-yl]benzenecarbonitrile 344-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-2-fluoro-benzonitrile 354-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indazol-5-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 364-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 374-[2-(4-aminopiperidyl)-1-methyl-5-(2-methyl (2H-indazol-5-yl))- A6-oxohydropyrimidin-4-yl]-2-fluorobenzenecarbonitrile 384-[2-(4-aminopiperidyl)-5-(3,5-difluoro-4-methoxyphenyl)-1-methyl- A6-oxohydropyrimidin-4-yl]benzenecarbonitrile 40{4-[2-(4-aminopiperidyl)-6-(4-cyanophenyl)-3-methyl-4-oxo B (3-hydropyrimidin-5-yl)]-2-fluorophenyl}-N-methylcarboxamide 414-[2-(4-aminopiperidyl)-6-(4-cyanophenyl)-3-methyl-4-oxo B (3-hydropyrimidin-5-yl)]-2-fluorobenzamide 424-[2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-(1-oxo-2,3-dihydro- B1H-isoindol-5-yl)-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 444-{5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3S)-(pyrrolidin- B3-ylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 454-{5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3R)-(pyrrolidin- B3-ylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 464-[2-[1,4]diazepan-1-yl-5-(3-fluoro-4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 472-fluoro-4-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo- B2-piperazin-1-yl-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 484-[5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-2-(piperidin- B4-ylamino)-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 494-[2-(4-amino-piperidin-1-yl)-2′-dimethylamino-1-methyl-6-oxo- A1,6-dihydro-[5,5′]bipyrimidinyl-4-yl]-2-fluoro-benzonitrile 505-[2-(4-amino-piperidin-1-yl)-4-(4-cyano-3-fluoro-phenyl)-1-methyl-6-oxo-A 1,6-dihydro-pyrimidin-5-yl]-pyridine-2-carboxylic acid methylamide 512-fluoro-4-{5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3S)-(pyrrolidin- B3-ylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 522-luoro-4-{5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-[(3R)-(pyrrolidin- B3-ylmethyl)-amino]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 532-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-6-oxo-2-(piperidin- B4-ylamino)-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 542-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-(3S)-pyrrolidin- A3-ylmethyl-amino)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 552-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-piperidin- B4-yl-amino)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 562-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(methyl-pyrrolidin- A3-ylmethyl-amino)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 574-[2-(4-amino-piperidin-1-yl)-5-(6-dimethylamino-pyridin-3-yl)-1-methyl-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 582-fluoro-4-[5-(6-methoxy-pyridin-3-yl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 594-[2-(4-amino-piperidin-1-yl)-5-(4-dimethylamino-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 604-[2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-(6-pyrrolidin-1-yl-pyridin-A 3-yl)-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 614-[2-[1,4]diazepan-1-yl-5-(6-methoxy-pyridin-3-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 624-[2-[1,4]diazepan-1-yl-5-(6-methoxy-pyridin-3-yl)-1-methyl- B6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 634-[2-[1,4]diazepan-1-yl-5-(6-dimethylamino-pyridin-3-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 644-[2-(3-amino-azetidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- B6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 652-fluoro-4-[1-methyl-2-(4-methylamino-piperidin-1-yl)-5-(2-methyl-2H- Aindazol-5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 664-[2-[1,4]diazepan-1-yl-1-methyl-5-(2-methyl-2H-indazol-5-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 674-[2-[1,4]diazepan-1-yl-5-(6-dimethylamino-pyridin-3-yl)-1- Amethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 684-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-morpholin-4-yl-pyridin-3-yl)-A 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 694-[2-(3-aminomethyl-azetidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- B6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 702-fluoro-4-[5-(4-methoxy-phenyl)-1-methyl-2-(3-methylaminomethyl- Aazetidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 714-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(2-methyl-2H-indazol- A5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 724-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indazol- A5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 734-[2-(4-amino-piperidin-1-yl)-5-(1H-indol-5-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 744-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indol-5-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 754-[2-(4-amino-piperidin-1-yl)-5-(1H-indol-6-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 764-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indol-6-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 774-[2-(4-amino-piperidin-1-yl)-5-(1H-indazol-6-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 784-[2-((4R,3S)-4-amino-3-fluoro-piperidin-1-yl)-5-(4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 794-[2-((4S,3R)-4-amino-3-fluoro-piperidin-1-yl)-5-(4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 804-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(2-methyl-2H-indazol- A6-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 814-[2′-dimethylamino-2-(4-dimethylamino-piperidin-1-yl)-1-methyl- B6-oxo-1,6-dihydro-[5,5′]bipyrimidinyl-4-yl]-2-fluoro-benzonitrile 824-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)-B 6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 834-[5-(6-dimethylamino-pyridin-3-yl)-1-methyl-2-(4-methylamino- Apiperidin-1-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile84 4-[2-(4-dimethylamino-piperidin-1-yl)-5-(2H-indazol-6-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 854-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1- Adeuteratedmethyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile864-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-deuteratedmethoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 872-fluoro-4-[1-methyl-2-[4-(methylamino)piperidin-1-yl]-5- A(1-methylindazol-5-yl)-6-oxopyrimidin-4-yl]benzonitrile 884-[2-(4-aminopiperidin-1-yl)-5-(1H-indazol-5-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 894-[5-(4-aminophenyl)-2-(4-aminopiperidin-1-yl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 904-[2-(4-aminopiperidin-1-yl)-1-methyl-5-[4-(methylamino)phenyl]- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 914-[2-(4-aminopiperidin-1-yl)-5-[3-fluoro-4-(methylamino)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 924-[2-[4-(dimethylamino)piperidin-1-yl]-5-(6-methoxypyridin-3-yl)- A1-methyl-6-oxo-pyrimidin-4-yl]-2-fluorobenzonitrile 934-[2-(4-aminopiperidin-1-yl)-5-(6-ethoxy-5-fluoropyridin-3-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 944-[2-(4-aminopiperidin-1-yl)-5-(6-ethoxypyridin-3-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 954-[2-(4-aminopiperidin-1-yl)-5-(4-ethoxyphenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 964-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethoxy)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 974-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethoxy)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]benzonitrile 984-[2-(4-aminopiperidin-1-yl)-5-[4-(2-methoxyethoxy)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 994-[2-(4-aminopiperidin-1-yl)-5-[4-(2-hydroxyethyl)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1004-[2-(4-aminopiperidin-1-yl)-5-[4-(hydroxymethyl)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1014-[2-(4-aminopiperidin-1-yl)-5-(4-fluorophenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1024-[2-(4-aminopiperidin-1-yl)-5-(3-fluorophenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1034-[2-(4-aminopiperidin-1-yl)-5-(3,5-difluorophenyl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1044-[2-(4-aminopiperidin-1-yl)-5-(3,4-difluorophenyl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1054-[2-(4-aminopiperidin-1-yl)-1-methyl-5-(4-methylsulfonyl Aphenyl)-6-oxo-pyrimidin-4-yl]-2-fluorobenzonitrile 1064-[2-(4-aminopiperidin-1-yl)-5-(4-chlorophenyl)-1-methyl- A6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1074-[2-(4-aminopiperidin-1-yl)-5-[4-(methoxymethyl)phenyl]- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile 1084-[2-(4-aminopiperidin-1-yl)-1-methyl-6-oxo- Bpyrimidin-4-yl]-2-fluorobenzonitrile 1104-[2-(4-amino-piperidin-1-yl)-1-cyclopropylmethyl-6-oxo- B1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 1112-(4-amino-piperidin-1-yl)-6-(4-chloro-3-fluoro-phenyl)-5- B(4-methoxy-phenyl)-3-methyl-3H-pyrimidin-4-one 1224-[2-(4-aminopiperidin-1-yl)-5-(4-methoxyphenyl)- A6-oxo-1H-pyrimidin-4-yl]-2-fluorobenzonitrile Note: Cellular assay IC₅₀data are designated within the following ranges: A: ≦0.10 μM; B: >0.10μM to ≦1.0 μM; C: >1.0 μM to ≦10 μM; D: >10 μM

Example 4: Kasumi-1 AML Cell Line Proliferation Assay (Cell-MTS Assay)

Colorimetric cellular assay to assess the ability of LSD-1 smallmolecule inhibitors to effect the proliferation of the established AMLcancer cell line Kasumi-1.

Assay Background

The LSD-1 protein has been shown to play a key role in the biology of avariety of cancer types including SCLC and AML. To demonstrate smallmolecule inhibition of LSD-1 as a potential anti-cancer therapy, anassay to measure the degree of proliferative inhibition in anestablished cancer cell line of AML was implemented.

Assay Principle

This Cell-MTS assay is a 7-day plate based colorimetric assay whichquantifies the amount of newly generated NADH in the presence andabsence of test compound. These NADH levels are used as a proxy for thequantification of cancer cell proliferation.

Assay Method

The established cancer cell line Kasumi-1 with a verified p53 mutationwere purchased from American Type Culture Collection (ATCC) androutinely passaged according to ATCC published protocols. For routineassay these cells were seeded at a density of 20,000 cells per 96-well.24 hours after plating, cells received an 11 point dilution of testcompound with final concentration ranges from 100 μM to 2.0 nM. Cellsare incubated in the presence of compound for 168 hours at 37° C., 5%CO₂. At the end of this compound incubation period, 80 μl of media isremoved and 20 μL of CellTiter 96® AQueous Non-Radioactive CellProliferation Assay solution (Promega) is added. The cells are incubateduntil the OD490 is >0.6. IC₅₀ values are calculated using the IDBS XLfitsoftware package and include background subtracted OD490 values andnormalization to DMSO controls.

Table 6 provides the Kasumi-1 cellular IC₅₀ values of varioussubstituted heterocyclic compounds disclosed herein.

TABLE 6 Chemical Synthesis Kasumi-1 Example Name IC₅₀ (μM) 14-(2-(4-aminopiperidin-1-yl)-1-methyl-6-oxo-5-p-tolyl- A1,6-dihydropyrimidin-4-yl)benzonitrile 34-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 44-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(6-methyl-pyridin-3-yl)- B6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 54-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 64-[2-(4-amino-piperidin-1-yl)-5-(4-methoxy-phenyl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 74-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)- A1-methyl-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 84-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 94-[2-(4-amino-piperidin-1-yl)-5-(6-methoxy-pyridin-3-yl)-1-methyl- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 244-[2-(4-aminopiperidyl)-1-methyl-5-(2-methyl(2H-indazol-5-yl))- A6-oxohydropyrimidin-4-yl]benzenecarbonitrile 344-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-2-fluoro-benzonitrile 354-[2-(4-amino-piperidin-1-yl)-1-methyl-5-(1-methyl-1H-indazol-5-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 364-{2-(4-amino-piperidin-1-yl)-1-methyl-6-oxo-5-[1-(2,2,2-trifluoro-ethyl)-A 1H-pyrazol-4-yl]-1,6-dihydro-pyrimidin-4-yl}-benzonitrile 652-fluoro-4-[1-methyl-2-(4-methylamino-piperidin-1-yl)-5-(2-methyl- A2H-indazol-5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-benzonitrile 664-[2-[1,4]diazepan-1-yl-1-methyl-5-(2-methyl-2H-indazol-5-yl)- A6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 714-[2-(4-dimethylamino-piperidin-1-yl)-1-methyl-5-(2-methyl-2H-indazol- A5-yl)-6-oxo-1,6-dihydro-pyrimidin-4-yl]-2-fluoro-benzonitrile 884-[2-(4-aminopiperidin-1-yl)-5-(1H-indazol-5-yl)- A1-methyl-6-oxopyrimidin-4-yl]-2-fluorobenzonitrile

Example 5: In Vivo Xenograph Study—MCF-7 Xenograph

Time release pellets containing 0.72 mg 17-3 Estradiol aresubcutaneously implanted into nu/nu mice. MCF-7 cells are grown in RPMIcontaining 10% FBS at 5% CO₂, 37° C. Cells are spun down andre-suspended in 50% RPMI (serum free) and 50% Matrigel at 1×10⁷cells/mL. MCF-7 cells are subcutaneously injected (100 L/animal) on theright flank 2-3 days post pellet implantation and tumor volume(length×width²/2) is monitored bi-weekly. When tumors reach an averagevolume of ˜200 mm³ animals are randomized and treatment is started.Animals are treated with vehicle or compound daily for 4 weeks. Tumorvolume and body weight are monitored bi-weekly throughout the study. Atthe conclusion of the treatment period, plasma and tumor samples aretaken for pharmacokinetic and pharmacodynamic analyses, respectively.

Example 6: In Vivo Xenograph Study—LNCaP Xenograph

LNCaP cells with a stable knockdown of LSDl (shLSDl cells) or controlcells (such as shNTC cells) are inoculated in the dorsal flank of nudemice by subcutaneous injection (such as 3×10⁶ cells in 100 μl of 50%RPMI 1640/BD Matrigel). Mouse weight and tumor size are measured onceper week and tumor volume is estimated using the formula (7i/6)(L×W),where L=length of tumor and W=width of tumor. A two sample t-test isperformed to determine statistical differences in mean tumor volumebetween the two groups.

Unmodified LNCaP cells are inoculated by subcutaneous injection into thedorsal flank of nude mice (such as 3×10⁶ cells in 100 μl of 50% RPMI1640/BD Matrigel). After three weeks, mice are injectedintraperitoneally once per day with water (control), pargyline (0.53 mgor 1.59 mg; 1 or 3 mM final concentration, assuming 70%bioavailability), or XB154 (4 or 20 μg; 1 or 5 μM final concentration,assuming 70% bioavailability) or treated with a test compound (5 mg/kgeach week or 10 mg/kg each week). Treatment continues for three weeks,during which time mouse weight and tumor volume are measured as above.

shLSDl LNCaP cells or control cells are injected in nude mice as above.After three weeks, mice are treated with 2.6 μg mitomycin C (predictedfinal concentration of 1 μM assuming 40% bioavailability), olaparib (forexample, about 0.5 mg/kg to 25 mg/kg), or vehicle intraperitoneally onceper day for three weeks. In other examples, unmodified LNCaP cells areinjected in nude mice as above.

After three weeks, mice are treated with test compounds, or vehicle asabove, plus MMC or olaparib. Treatment continues for three weeks, duringwhich time mouse weight and tumor volume are measured as above.

A decrease in tumor volume compared to control in mice injected withshLSDl cells indicates that LSDl inhibition decreases tumor growth invivo.

Similarly, a decrease in tumor volume compared to control in miceinjected with LNCaP cells and treated with a compound disclosed hereinindicates that LSDl inhibition decreases tumor growth in vivo. Finally,a decrease in tumor volume in mice injected with LNCaP cells and treatedwith a compound disclosed herein plus olaparib as compared to micetreated with a compound disclosed herein alone indicates that inhibitionof LSD1 plus inhibition of PARP decreases tumor growth in vivo.

The harvested xenograft tissue is examined for evidence of LSD1inhibition. This is assessed with Western blots to examine global levelsof the 2MK4 and 2MK9 histone marks, expression of FA/BRCA genes, FANCD2ubiquitination, and LSD1 protein levels in the cases of the shRNA cells.A decrease in one or more of these parameters indicates the effectiveinhibition of LSD 1. Additionally, effects on DNA damage repair areassessed with staining for H2AX foci.

III. Preparation of Pharmaceutical Dosage Forms Example 1: Oral Tablet

A tablet is prepared by mixing 48% by weight of a compound of Formula(I), or a pharmaceutically acceptable salt thereof, 45% by weight ofmicrocrystalline cellulose, 5% by weight of low-substitutedhydroxypropyl cellulose, and 2% by weight of magnesium stearate. Tabletsare prepared by direct compression. The total weight of the compressedtablets is maintained at 250-500 mg.

We claim:
 1. A compound, or pharmaceutically acceptable salt thereof,chosen from:


2. A pharmaceutical composition comprising a pharmaceutically acceptablecarrier and a compound of claim 1, or a pharmaceutically acceptable saltthereof.